Tau-conotoxin peptides

ABSTRACT

The invention relates to relatively short peptides (termed τ-conotoxins herein), about 10-25 residues in length, which are naturally available in minute amounts in the venom of the cone snails or analogous to the naturally available peptides, and which preferably include two disulfide bonds.

CROSS-REFERENCE TO RELATED APPLICATION

The present application is related to U.S. provisional patentapplication Ser. No. 60/118,642 filed on Feb. 4, 1999, incorporatedherein by reference.

This invention was made with Government support under Grant No. PO 1GM48677 awarded by the National Institute of General Medical Sciences,National Institutes of Health, Bethesda, Md. The United StatesGovernment has certain rights in the invention.

BACKGROUND OF THE INVENTION

The invention relates to relatively short peptides (termed τ-conotoxinsherein), about 10-20 residues in length, which are naturally availablein minute amounts in the venom of the cone snails or analogous to thenaturally available peptides, and which preferably include two disulfidebonds.

The publications and other materials used herein to illuminate thebackground of the invention, and in particular, cases to provideadditional details respecting the practice, are incorporated byreference, and for convenience are referenced in the following text byauthor and date and are listed alphabetically by author in the appendedbibliography.

The predatory cone snails (Conus) have developed a unique biologicalstrategy. Their venom contains relatively small peptides that aretargeted to various neuromuscular receptors and may be equivalent intheir pharmacological diversity to the alkaloids of plants or secondarymetabolites of microorganisms. Many of these peptides are among thesmallest nucleic acid-encoded translation products having definedconformations, and as such, they are somewhat unusual. Peptides in thissize range normally equilibrate among many conformations. Proteinshaving a fixed conformation are generally much larger.

The cone snails that produce these peptides are a large genus ofvenomous gastropods comprising approximately 500 species. All cone snailspecies are predators that inject venom to capture prey, and thespectrum of animals that the genus as a whole can envenomate is broad. Awide variety of hunting strategies are used, however, every Conusspecies uses fundamentally the same basic pattern of envenomation.

Several peptides isolated from Conus venoms have been characterized.These include the α-, μ- and ω-conotoxins which target nicotinicacetylcholine receptors, muscle sodium channels, and neuronal calciumchannels, respectively (Olivera et at., 1985). Conopressins, which arevasopressin analogs, have also been identified (Cruz et al. 1987). Inaddition, peptides named conantokins have been isolated from Conusgeographus and Conus tulipa (Mena et al., 1990; Haack et al., 1990).

Chronic or intractable pain, which may result from degenerativeconditions or debilitating diseases, is currently treated with a varietyof analgesic compounds, often opioid compounds such as morphine.Likewise, neuropathic pain, typically a chronic condition attributableto injury or partial transection of a peripheral nerve, is alsoconventionally treated with opioid compounds such as morphine.

Conventional therapies for pain produce analgesia—a loss of sensitivityto pain without the loss of consciousness. Opioid compounds have beenused widely to produce analgesia, including plant-derived opioids suchas morphine, and endogenous opioids such as met- and leu-enkephalins, aswell as beta-endorphin.

Opioid compounds, while effective in producing analgesia for many typesof pain, may induce tolerance in some patients. When a patient becomestolerant, increasing doses of the opioid are required to produce thedesired analgesic effect. In addition, these compounds frequently resultin a physical dependence in patients, and may have side effects at highdoses.

The analgesic effects and adverse actions of variousN-methyl-D-aspartate (NMDA) receptor antagonists has been shown to varydepending on the site of action and potency of the drug. For example,NMDA receptor antagonists acting at the ion channel in a noncompetitivemanner (e.g., MK-801 and phenylcyclidine (PCP)) or competitiveinhibitors, show analgesic activity but show motor impairment atequivalent doses. Glycine B-site NMDA antagonists appear to haveanalgesic activity at doses that do not impair motor function.Conantokins, which are polyamine-site NMDA antagonist compounds haveanalgesic effects at doses which do not produce overt side effects (PCTpublished application WO 98/03189).

It is desired to provide additional compounds which have analgesicproperties.

SUMMARY OF THE INVENTION

The invention relates to relatively short peptides (termed τ-conotoxinsherein), about 10-25 residues in length, which are naturally availablein minute amounts in the venom of the cone snails or analogous to thenaturally available peptides, and which preferably include two disulfidebonds.

More specifically, the present invention is directed to τ-conotoxinpeptides having the general formula I:

Xaa₁-Xaa₂-Xaa₃-Xaa₄-Cys-Cys-Xaa₅-Xaa₆-Xaa₇-Xaa₈-Xaa₉-Cys-Cys-Xaa₁₀-Xaa₁₁-Xaa₁₂-Xaa₁₃-Xaa₁₄-Xaa₁₅-Xaa₁₆-Xaa₁₇-Xaa₁₈-Xaa₁₉(SEQ ID NO:1), wherein Xaa₁ is des-Xaa₁, Asp, Glu or γ-carboxy-Glu(Gla); Xaa₂ is des-Xaa₂, Gln, Asn, Glu, Trp (D or L), neo-Trp, halo-Trpor any unnatural aromatic amino acid; Xaa₃ is des-Xaa₃, Gly, Ala, Asn orGln; Xaa₄ is des-Xaa₄, Val, Leu (D or L), Ile, Ala, Gly, Glu, Gla, Asp,Ser, Thr, Phe, Trp (D or L), neo-Trp, halo-Trp (D or L) or any unnaturalaromatic amino acid; Xaa₅ is Pro, hydroxy-Pro, Gln, Asn, Glu, Gla, Ala,Gly, Lys, Arg, Ile, Val, homoarginine, ornithine, N-methyl-Lys,N,N-dimethyl-Lys, N,N,N-trimethyl-Lys or any unnatural basic amino acid;Xaa₆ is Val, Phe, Thr, Ser, Glu, Gla, Asp, Asn, Gln, Ala, Gly, Ile, Leu(D or L) Met, Pro, hydroxy-Pro, Arg, homoarginine, ornithine, Lys,N-methyl-Lys, N,N-dimethyl-Lys, N,N,N-trimethyl-Lys, any unnatural basicamino acid or any unnatural aromatic amino acid; Xaa₇ is any Val, Ile,Asn, Leu (D or L), Gln, Gly, Ala, Phe, Glu, Gla, Arg, ornithine,homoarginine, Lys, N-methy-Lys, N,N-dimethyl-Lys, N,N,N-trimethyl-Lys,any unnatural basic amino acid or any unnatural aromatic amino acid;Xaa₈ is Ile, Leu (D or L), Met, Thr, Ser, Pro, hydroxy-Pro, Gln, Asp,Glu, Gla, Asn, Arg, homoarginine, ornithine, Lys, N-methy-Lys,N,N-dimethyl-Lys, N,N,N-trimethyl-Lys, Tyr, nor-Tyr, mono-halo-Tyr,di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr, nitro-Tyr, any unnatural basicamino acid, any unnatural aromatic amino acid or any unnatural hydroxycontaining amino acid; Xaa₁ is des-Xaa₉, Ala, Gly, Asp, Glu, Gla, Trp (Dor L) neo-Trp, halo-Trp (D or L), Lys, N-methy-Lys, N,N-dimethyl-Lys,N,N,N-trimethyl-Lys, Arg, homoarginine, ornithine, Tyr, nor-Tyr,mono-halo-Tyr, di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr, nitro-Tyr orany unnatural basic amino acid; Xaa₁₀ is des-Xaa₁₀, Ile, Leu (D or L),Val, Glu, Gla, Asp, Thr, Ser, Pro, hydroxy-Pro, Trp (D or L), neo-Trp,halo-Trp (D or L), Phe, any unnatural aromatic amino acid or anyunnatural hydroxy containing amino acid; Xaa₁, is des-Xaa₁₁, Gln, Asn,Leu (D or L), Ile, Val, Ala, Gly, Trp (D or L), neo-Trp, halo-Trp (D orL), Arg, homoarginine, ornithine, Lys, N-methy-Lys, N,N-dimethyl-Lys,N,N,N-trimethyl-Lys, any unnatural basic amino acid or any unnaturalaromatic amino acid; Xaa₁₂ is des-Xaa₁₂, Ala, Gly, Phe, Trp (D or L),neo-Trp, halo-Trp (D or L) or any unnatural aromatic amino acid; Xaa₁₃is des-Xaa₁₃, Glu, Gla, Asp, Phe or any unnatural aromatic amino acid;Xaa₁₄ is des-Xaa₁₄, Ile, Val or Leu (D or L); Xaa₁₅ is des-Xaa₁₅, Thr,Ser, Arg, homoarginine, ornithine, Lys, N-methy-Lys, N,N-dimethyl-Lys,N,N,N-trimethyl-Lys or any unnatural basic amino acid; Xaa₁₆ isdes-Xaa₁₆, Glu, Gla or Asp; Xaa₁₇ is des-Xaa₁₇, Asn or Gln; Xaa₁₈ isdes-Xaa₁₈, Asp, Glu or Gla; Xaa₁₉ is des-Xaa₁₉, Phe or any unnaturalaromatic amino acid. The C-terminus may contain a free carboxyl group oran amide group. The halo is preferably bromine, chlorine or iodine, morepreferably iodine for Tyr and bromine for Trp. The Cys residues may bein D or L configuration and may optionally be substituted withhomocysteine (D or L). The Tyr residues may be substituted with the3-hydroxyl or 2-hydroxyl isomers and corresponding O-sulpho- andO-phospho-derivatives. The acidic amino acid residues may be substitutedwith any synthetic acidic amino acid, e.g., tetrazolyl derivatives ofGly and Ala.

The present invention is also directed to novel specific τ-conotoxinpeptides of general formula I having the formulas:

Phe-Cys-Cys-Xaa₁-Val-Ile-Arg-Xaa₂-Cys-Cys-Xaa₃ (SEQ ID NO:2);

Phe-Cys-Cys-Xaa₁-Phe-Ile-Arg-Xaa₂-Cys-Cys-Xaa₃ (SEQ ID NO:3);

Cys-Cys-Gln-Thr-Phe-Xaa₂-Xaa₃-Cys-Cys-Gln (SEQ ID NO:4);

Xaa₄-Gly-Xaa₃-Cys-Cys-Xaa₅-Xaa₆-Asn-Ile-Ala-Cys-Cys-Ile (SEQ ID NO:5);

Gly-Cys-Cys-Ala-Arg-Leu-Thr-Cys-Cys-Val (SEQ ID NO:6);

Asn-Gly-Cys-Cys-Xaa₁-Xaa₅-Gln-Met-Arg-Cys-Cys-Thr (SEQ ID NO:7);

Asp-Xaa₃-Asn-Ser-Cys-Cys-Gly-Xaa₆-Asn-Xaa₁-Gly-Cys-Cys-Xaa₁-Xaa₃ (SEQ IDNO:8);

Xaa₄-Gly-Xaa₃-Cys-Cys-Xaa₅-Xaa₆-Asn-Ile-Arg-Cys-Cys-Val (SEQ ID NO:9);

Xaa₆-Cys-Cys-Xaa₆-Asp-Gly-Xaa₃-Cys-Cys-Thr-Ala-Ala-Xaa₁-Leu-Thr (SEQ IDNO:10);

Gly-Cys-Cys-Xaa₆-Asp-Gly-Xaa₃-Cys-Cys-Thr-Ala-Ala-Xaa₁-Leu-Thr(SEQ IDNO:11);

Asn-Gly-Cys-Cys-Arg-Ala-Gly-Asp-Cys-Cys-Ser-Arg-Phe-Xaa₆-Ile-Xaa₅-Xaa₆-Asn-Asp-Phe(SEQ ID NO:12);

Asn-Ala-Cys-Cys-Ile-Val-Arg-Gln-Cys-Cys (SEQ ID NO:13);

Asn-Gly-Cys-Cys-Arg-Ala-Gly-Asp-Cys-Cys-Ser (SEQ ID NO:14);

Cys-Cys-Xaa₁-Arg-Arg-Leu-Ala-Cys-Cys-Ile-lie (SEQ ID NO:15);

Cys-Cys-Xaa₁-Asn-Xaa₅-Xaa₁-Cys-Cys-Phe-Ile (SEQ ID NO:16);

Gly-Cys-Cys-Ala-Met-Leu-Thr-Cys-Cys-Val (SEQ ID NO:17);

Leu-Cys-Cys-Val-Thr-Xaa₆-Asp-Xaa₃-Cys-Cys-Xaa₆-Xaa₃-Xaa₃ (SEQ ID NO:18);and

Val-Cys-Cys-Arg-Xaa₁-Val-Gln-Asp-Cys-Cys-Ser (SEQ ID NO:19);

wherein Xaa₁ is Pro or hydroxy-Pro; Xaa₂ is Tyr, mono-halo-Tyr,di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr or nitro-Tyr; Xaa₃ is Trp orhalo-Trp; Xaa₄ is Gln or pyro-Glu; Xaa₅ is Lys, N-methyl-Lys,N,N-dimethyl-Lys or N,n,N-trimethyl-Lys, Xaa₆ is Glu orgamma-carboxy-Glu (Gla); and the C-terminus contains a carboxyl or amidegroup. The halo is preferably bromine, chlorine or iodine, morepreferably iodine for Tyr and bromine for Trp. In addition, the Argresidues may be substituted by Lys, ornithine, homoargine, N-methyl-Lys,N,N-dimethyl-Lys, N,N,N-trimethyl-Lys or any unnatural basic amino acid;the Lys residues may be substituted by Arg, ornithine, homoargine,N-methyl-Lys, N,N-dimethyl-Lys, N,N,N-trimethyl-Lys or any unnaturalbasic amino acid; the Tyr residues may be substituted with any unnaturalhydroxy containing amino acid; the Ser residues may be substituted withThr; the Thr residues may be substituted with Ser; and the Phe and Trpresidues may be substituted with any unnatural aromatic amino acid. TheCys residues may be in D or L configuration and may optionally besubstituted with homocysteine (D or L). The Tyr residues may besubstituted with the 3-hydroxyl or 2-hydroxyl isomers and correspondingO-sulpho- and O-phospho-derivatives. The acidic amino acid residues maybe substituted with any synthetic acidic amino acid, e.g., tetrazolylderivatives of Gly and Ala.

More specifically, the present invention is directed to the followingτ-conotoxin peptides of general formula I:

AuVA: SEQ ID NO:2, wherein Xaa₁ is Pro, Xaa₂ is Tyr and Xaa₃ is Trp;

AuVB: SEQ ID NO:3, wherein Xaa₁ is Pro, Xaa₂ is Tyr and Xaa₃ is Trp;

Tx5.1: SEQ ID NO:4, wherein Xaa₂ is Tyr and Xaa₃ is Trp;

G5.1: SEQ ID NO:5, wherein Xaa₃ is Trp, Xaa₄ is Gln, Xaa₅ is Lys andXaa₆is Glu;

Qc5.1: SEQ ID NO:6;

PVA: SEQ ID NO:7, wherein Xaa₁ is Pro and Xaa₅ is Lys;

Im5.1: SEQ ID NO:8, wherein Xaa₁ is Pro, Xaa₃ is Trp and Xaa₁ is Lys;

G5.2: SEQ ID NO:9, wherein Xaa₃ is Trp, Xaa₄ is Gln, Xaa₅ is Lys andXaa₆ is Glu;

Tx5.2a: SEQ ID NO:10, wherein Xaa₁ is Pro, Xaa₃ is Trp and Xaa₆ is Glu;

Tx5.2b: SEQ ID NO:11, wherein Xaa₁ is Pro, Xaa₃ is Trp and Xaa₆ is Glu;

Mr5.1: SEQ ID NO:12, wherein Xaa₁ is Lys and Xaa₆ is Glu;

Mr5.2: SEQ ID NO:13;

Mr5.3: SEQ ID NO:14;

Ca5.1: SEQ ID NO:15, wherein Xaa₁ is Pro;

Ca5.2: SEQ ID NO:16, wherein Xaa₁ is Pro and Xaa₅ is Lys;

Qc5.2: SEQ ID NO:17;

Gm5.1: SEQ ID NO:18, wherein Xaa₃ is Trp and Xaa₆ is Glu; and

Gm5.2: SEQ ID NO:19, wherein Xaa₁ is Pro.

The C-terminus preferably contains a carboxyl group for the peptidesAuVA, AuVB, G5.1, PVA, G5.2, Mr5.2, Mr5.3 and Gm5.1 The C-terminus ofthe other peptides preferably contains an amide group.

Examples of unnatural aromatic amino acid include, but are not limitedto, such as nitro-Phe, 4-substituted-Phe wherein the substituent isC₁-C₃ alkyl, carboxyl, hyrdroxymethyl, sulphomethyl, halo, phenyl, —CHO,—CN, —SO₃H and —NHAc. Examples of unnatural hydroxy containing aminoacid, include, but are not limited to, such as 4-hydroxymethyl-Phe,4-hydroxyphenyl-Gly, 2,6-dimethyl-Tyr and 5-amino-Tyr. Examples ofunnatural basic amino acids include, but are not limited to,N-1-(2-pyrazolinyl)-Arg, 2-(4-piperinyl)-Gly, 2-(4-piperinyl)-Ala,2-[3-(2S)pyrrolininyl)-Gly and 2-[3-(2S)pyrrolininyl)-Ala. These andother unnatural basic amino acids, unnatural hydroxy containing aminoacids or unnatural aromatic amino acids are described in Building BlockIndex, Version 3.0 (1999 Catalog, pages 4-47 for hydroxy containingamino acids and aromatic amino acids and pages 66-87 for basic aminoacids), incorporated herein by reference, by and available from RSPAmino Acid Analogues, Inc., Worcester, Mass. Examples of synthetic acidamino acids include those derivatives bearing acidic functionality,including carboxyl, phosphate, sulfonate and synthetic tetrazolylderivatives such as described by Ornstein et al. (1993) and in U.S. Pat.No. 5,331,001, each incorporated herein by reference.

Optionally, in the peptides of general formula I and the specificpeptides described above, the Asn residues may be modified to contain anN-glycan and the Ser and Thr residues may be modified to contain anO-glycan. In accordance with the present invention, a glycan shall meanany N-, S- or O-linked mono-, di-, tri-, poly- or oligosaccharide thatcan be attached to any hydroxy, amino or thiol group of natural ormodified amino acids by synthetic or enzymatic methodologies known inthe art. The monosaccharides making up the glycan can include D-allose,D-altrose, D-glucose, D-mannose, D-gulose, D-idose, D-galactose,D-talose, D-galactosamine, D-glucosamine, D-N-acetyl-glucosamine(GIcNAc), D-N-acetyl-galactosamine (GalNAc), D-fucose or D-arabinose.These saccharides may be structurally modified, e.g., with one or moreO-sulfate, O-phosphate, O-acetyl or acidic groups, such as sialic acid,including combinations thereof. The glycan may also include similarpolyhydroxy groups, such as D-penicillamine 2,5 and halogenatedderivatives thereof or polypropylene glycol derivatives. The glycosidiclinkage is beta and 1-4 or 1-3, preferably 1-3. The linkage between theglycan and the amino acid may be alpha or beta, preferably alpha and is1−.

Core O-glycans have been described by Van de Steen et al. (1998),incorporated herein by reference. Mucin type O-linked oligosaccharidesare attached to Ser or Thr (or other hydroxylated residues of thepresent peptides) by a GalNAc residue. The monosaccharide buildingblocks and the linkage attached to this first GalNAc residue define the“core glycans,” of which eight have been identified. The type ofglycosidic linkage (orientation and connectivities) are defined for eachcore glycan. Suitable glycans and glycan analogs are described furtherin U.S. patent application Ser. No. 09/420,797, filed Oct. 19 , 1999 andin PCT Application No. PCT/US99/24380, filed Oct. 19, 1999, bothincorporated herein by reference. A preferred glycan isGal(β1→3)GalNAc(α1→).

Optionally, in the peptides of general formulas I and II and thespecific peptides described above, pairs of Cys residues may be replacedpairwise with isosteric lactam or ester-thioether replacements, such asSer/(Glu or Asp), Lys/(Glu or Asp) or Cys/Ala combinations. Sequentialcoupling by known methods (Barnay et al., 2000; Hruby et al., 1994;Bitan et al., 1997) allows replacement of native Cys bridges with lactambridges. Thioether analogs may be readily synthesized using halo-Alaresidues commercially available from RSP Amino Acid Analogues.

The present invention is further directed to propeptides and nucleicacid sequences encoding the propeptides or peptides as described infurther detail herein.

SUMMARY OF THE SEQUENCE LISTING

SEQ ID NO:1is generic formula I for τ-conotoxin peptides. SEQ ID NO:2 isa generic formula for the peptide AuVA. SEQ ID NO:3 is a generic formulafor the peptide AuVB. SEQ ID NO:4 is a generic formula for the peptideTx5.1. SEQ ID NO:5 is a generic formula for the peptide G5.1. SEQ IDNO:6 is a generic formula for the peptide Qc5.1. SEQ ID NO:7 is ageneric formula for the peptide PVA. SEQ ID NO:8 is a generic formulafor the peptide Im5.1. SEQ ID NO:9 is a generic sequence for the peptideG5.2. SEQ ID NO:10 is a generic sequence for the peptide Tx5.2a. SEQ IDNO:11 is a generic sequence for the peptide Tx5.2b. SEQ ID NO:12 is ageneric sequence for the peptide Mr5.1. SEQ ID NO:13 is a genericsequence for the peptide Mr5.2. SEQ ID NO:14 is a generic formula forthe peptide Mr5.3. SEQ ID NO:15 is a generic formula for the peptideCa5.1. SEQ ID NO:16 is a generic formula for the peptide Ca5.2. SEQ IDNO:17 is a generic formula for the peptide Qc5.2. SEQ ID NO:18 is ageneric formula for the peptide Gm5.1. SEQ ID NO:19 is a generic formulafor the peptide Gm5.2. SEQ ID NO:20 is a DNA sequence coding for theTx5.1 propeptide. SEQ ID NO:21 is the amino acid sequence of the Tx5.1propeptide. SEQ ID NO:22 is a DNA sequence coding for the G5.1propeptide. SEQ ID NO:23 is the amino acid sequence of the G5.1propeptide. SEQ ID NO:24 is a DNA sequence coding for the Qc5.1propeptide. SEQ ID NO:25 is the amino acid sequence of the Qc5.1propeptide. SEQ ID NO:26 is a DNA sequence coding for the Im5.1propeptide. SEQ ID NO:27 is the amino acid sequence of the Im5.1propeptide. SEQ ID NO:28 is a DNA sequence coding for the G5.2propeptide. SEQ ID NO:29 is the amino acid sequence of the G5.2propeptide. SEQ ID NO:30 is a DNA sequence coding for the Tx5.2propeptide. SEQ ID NO:31 is the amino acid sequence of the Tx5.2propeptide. SEQ ID NO:32 is a DNA sequence coding for the Tx5.3propeptide. SEQ ID NO:33 is the amino acid sequence of the Tx5.3propeptide. SEQ ID NO:34 is a DNA sequence coding for the Mr5.1 peptide.SEQ ID NO:35 is the amino acid sequence of the Mr5.1 peptide. SEQ IDNO:36 is a DNA sequence coding for the Mr5.2 peptide. SEQ ID NO:37 isthe amino acid sequence of the Mr5.2 peptide. SEQ ID NO:38 is a DNAsequence coding for the Mr5.3 propeptide. SEQ ID NO:39 is the amino acidsequence of the Mr5.3 propeptide. SEQ ID NO:40 is a DNA sequence codingfor the Ca5.1 propeptide. SEQ ID NO:41 is the amino acid sequence of theCa5.1 propeptide. SEQ ID NO:42 is a DNA sequence coding for the Ca5.2propeptide. SEQ ID NO:43 is the amino acid sequence of the Ca5.2propeptide. SEQ ID NO:44 is a DNA sequence coding for the Qc5.2propeptide. SEQ ID NO:45 is the amino acid sequence of the Qc5.2propeptide. SEQ ID NO:46 is a DNA sequence coding for the Gm5.1propeptide. SEQ ID NO:47 is the amino acid sequence of the Gm5.1propeptide. SEQ ID NO:48 is a DNA sequence coding for the Gm5.2propeptide. SEQ ID NO:49 is the amino acid sequence of the Gm5.2propeptide.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

The invention relates to relatively short peptides (termed r-conotoxinsherein), about 10-25 residues in length, which are naturally availablein minute amounts in the venom of the cone snails or analogous to thenaturally available peptides, and which preferably include two disulfidebonds.

The present invention, in another aspect, relates to a pharmaceuticalcomposition comprising an effective amount of an τ-conotoxin peptide, amutein thereof, an analog thereof, an active fragment thereof orpharmaceutically acceptable salts . Such a pharmaceutical compositionhas the capability of acting as an antagonist for acetylcholinereceptors and as analgesic agents for the treatment of pain, includingmigraine. Thus, the pharmaceutical compositions of the present inventionare useful in the treatment of pain (whether acute or chronic),including chronic pain, and neuropathic pain, without undesirable sideeffects.

The τ-conotoxin peptides described herein are sufficiently small to bechemically synthesized. General chemical syntheses for preparing theforegoing τ-conotoxin peptides are described hereinafter. Various onesof the τ-conotoxin peptides can also be obtained by isolation andpurification from specific Conus species using the technique describedin U.S. Pat. Nos. 4,447,356 (Olivera et al., 1984); 5,514,774;5,719,264; and 5,591,821, as well as in PCT published application WO98/03189, the disclosures of which are incorporated herein by reference.

Although the τ-conotoxin peptides of the present invention can beobtained by purification from cone snails, because the amounts ofτ-conotoxin peptides obtainable from individual snails are very small,the desired substantially pure τ-conotoxin peptides are best practicallyobtained in commercially valuable amounts by chemical synthesis usingsolid-phase strategy. For example, the yield from a single cone snailmay be about 10 micrograms or less of τ-conotoxin peptide. By“substantially pure” is meant that the peptide is present in thesubstantial absence of other biological molecules of the same type; itis preferably present in an amount of at least about 85% purity andpreferably at least about 95% purity. Chemical synthesis of biologicallyactive τ-conotoxin peptides depends of course upon correct determinationof the amino acid sequence.

The τ-conotoxin peptides can also be produced by recombinant DNAtechniques well known in the art. Such techniques are described bySambrook et al. (1989). A gene of interest (i.e., a gene that encodes asuitable τ-conotoxin peptide) can be inserted into a cloning site of asuitable expression vector by using standard techniques. Thesetechniques are well known to those skilled in the art. The expressionvector containing the gene of interest may then be used to transfect thedesired cell line. Standard transfection techniques such as calciumphosphate co-precipitation, DEAE-dextran transfection or electroporationmay be utilized. A wide variety of host/expression vector combinationsmay be used to express a gene encoding a conotoxin peptide of interest.Such combinations are well known to a skilled artisan. The peptidesproduced in this manner are isolated, reduced if necessary, and oxidizedto form the correct disulfide bonds.

One method of forming disulfide bonds in the τ-conotoxin peptides of thepresent invention is the air oxidation of the linear peptides forprolonged periods under cold room temperatures or at room temperature.This procedure results in the creation of a substantial amount of thebioactive, disulfide-linked peptides. The oxidized peptides arefractionated using reverse-phase high performance liquid chromatography(HPLC) or the like, to separate peptides having different linkedconfigurations. Thereafter, either by comparing these fractions with theelution of the native material or by using a simple assay, theparticular fraction having the correct linkage for maximum biologicalpotency is easily determined. However, because of the dilution resultingfrom the presence of other fractions of less biopotency, a somewhathigher dosage may be required.

The peptides are synthesized by a suitable method, such as byexclusively solid-phase techniques, by partial solid-phase techniques,by fragment condensation or by classical solution couplings.

In conventional solution phase peptide synthesis, the peptide chain canbe prepared by a series of coupling reactions in which constituent aminoacids are added to the growing peptide chain in the desired sequence.Use of various coupling reagents, e.g., dicyclohexylcarbodiimide ordiisopropylcarbonyldimidazole, various active esters, e.g., esters ofN-hydroxyphthalimide or N-hydroxy-succinimide, and the various cleavagereagents, to carry out reaction in solution, with subsequent isolationand purification of intermediates, is well known classical peptidemethodology. Classical solution synthesis is described in detail in thetreatise, “Methoden der Organischen Chemie (Houben-Weyl): Synthese vonPeptiden,” (1974). Techniques of exclusively solid-phase synthesis areset forth in the textbook, “Solid-Phase Peptide Synthesis,” (Stewart andYoung, 1969), and are exemplified by the disclosure of U.S. Pat. No.4,105,603 (Vale et al., 1978). The fragment condensation method ofsynthesis is exemplified in U.S. Pat. No. 3,972,859 (1976). Otheravailable syntheses are exemplified by U.S. Pat. Nos. 3,842,067 (1974)and 3,862,925 (1975). The synthesis of peptides containingγ-carboxyglutamic acid residues is exemplified by Rivier et al. (1987),Nishiuchi et al. (1993) and Zhou et al. (1996).

Common to such chemical syntheses is the protection of the labile sidechain groups of the various amino acid moieties with suitable protectinggroups which will prevent a chemical reaction from occurring at thatsite until the group is ultimately removed. Usually also common is theprotection of an α-amino group on an amino acid or a fragment while thatentity reacts at the carboxyl group, followed by the selective removalof the α-amino protecting group to allow subsequent reaction to takeplace at that location. Accordingly, it is common that, as a step insuch a synthesis, an intermediate compound is produced which includeseach of the amino acid residues located in its desired sequence in thepeptide chain with appropriate side-chain protecting groups linked tovarious ones of the residues having labile side chains.

As far as the selection of a side chain amino protecting group isconcerned, generally one is chosen which is not removed duringdeprotection of the α-amino groups during the synthesis. However, forsome amino acids, e.g., His, protection is not generally necessary. Inselecting a particular side chain protecting group to be used in thesynthesis of the peptides, the following general rules are followed: (a)the protecting group preferably retains its protecting properties and isnot split off under coupling conditions, (b) the protecting group shouldbe stable under the reaction conditions selected for removing theα-amino protecting group at each step of the synthesis, and (c) the sidechain protecting group must be removable, upon the completion of thesynthesis containing the desired amino acid sequence, under reactionconditions that will not undesirably alter the peptide chain.

It should be possible to prepare many, or even all, of these peptidesusing recombinant DNA technology. However, when peptides are not soprepared, they are preferably prepared using the Merrifield solid-phasesynthesis, although other equivalent chemical syntheses known in the artcan also be used as previously mentioned. Solid-phase synthesis iscommenced from the C-terminus of the peptide by coupling a protectedα-amino acid to a suitable resin. Such a starting material can beprepared by attaching an α-amino-protected amino acid by an esterlinkage to a chloromethylated resin or a hydroxymethyl resin, or by anamide bond to a benzhydrylamine (BHA) resin or paramethylbenzhydrylamine(MBHA) resin. Preparation of the hydroxymethyl resin is described byBodansky et al. (1966). Chloromethylated resins are commerciallyavailable from Bio Rad Laboratories (Richmond, Calif.) and from Lab.Systems, Inc. The preparation of such a resin is described by Stewartand Young (1969). BHA and MBHA resin supports are commerciallyavailable, and are generally used when the desired polypeptide beingsynthesized has an unsubstituted amide at the C-terminus. Thus, solidresin supports may be any of those known in the art, such as one havingthe formulae —O—CH₂-resin support, —NH BHA resin support, or —NH—MBHAresin support. When the unsubstituted amide is desired, use of a BHA orMBHA resin-is preferred, because cleavage directly gives the amide. Incase the N-methyl amide is desired, it can be generated from an N-methylBHA resin. Should other substituted amides be desired, the teaching ofU.S. Pat. No. 4,569,967 (Kornreich et al., 1986) can be used, or shouldstill other groups than the free acid be desired at the C-terminus, itmay be preferable to synthesize the peptide using classical methods asset forth in the Houben-Weyl text (1974).

The C-terminal amino acid, protected by Boc or Fmoc and by a side-chainprotecting group, if appropriate, can be first coupled to achloromethylated resin according to the procedure set forth in K. Horikiet al. (1978), using KF in DMF at about 60° C. for 24 hours withstirring, when a peptide having free acid at the C-terminus is to besynthesized. Following the coupling of the BOC-protected amino acid tothe resin support, the α-amino protecting group is removed, as by usingtrifluoroacetic acid (TFA) in methylene chloride or TFA alone. Thedeprotection is carried out at a temperature between about 0° C. androom temperature. Other standard cleaving reagents, such as HCl indioxane, and conditions for removal of specific α-amino protectinggroups may be used as described in Schroder & Lubke (1965).

After removal of the α-amino-protecting group, the remaining α-amino-and side chain-protected amino acids are coupled step-wise in thedesired order to obtain the intermediate compound defined hereinbefore,or as an alternative to adding each amino acid separately in thesynthesis, some of them may be coupled to one another prior to additionto the solid phase reactor. Selection of an appropriate coupling reagentis within the skill of the art. Particularly suitable as a couplingreagent is N,N′-dicyclohexylcarbodiimide (dicyclohexylcarbodiimide(DCC),diisopropylcarbodiimide (DIC),O-(benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate(HBTU), O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (HATU),O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate(TBTU) in the presence of HoBt or HoAt).

The activating reagents used in the solid phase synthesis of thepeptides are well known in the peptide art. Examples of suitableactivating reagents are carbodiimides, such asN,N′-diisopropylcarbodiimide andN-ethyl-N′-(3-dimethylaminopropyl)carbodiimide. Other activatingreagents and their use in peptide coupling are described by Schroder &Lubke (1965) and Kapoor (1970).

Each protected amino acid or amino acid sequence is introduced into thesolid-phase reactor in about a twofold or more excess, and the couplingmay be carried out in a medium of dimethylformamide (DMF):CH₂Cl₂ (1:1)or in DMF or CH₂Cl₂ alone. In cases where intermediate coupling occurs,the coupling procedure is repeated before removal of the α-aminoprotecting group prior to the coupling of the next amino acid. Thesuccess of the coupling reaction at each stage of the synthesis, ifperformed manually, is preferably monitored by the ninhydrin reaction,as described by Kaiser et al. (1970). Coupling reactions can beperformed automatically, as on a Beckman 990 automatic synthesizer,using a program such as that reported in Rivier et al. (1978).

After the desired amino acid sequence has been completed, theintermediate peptide can be removed from the resin support by treatmentwith a reagent, such as liquid hydrogen fluoride or TFA (if using Fmocchemistry), which not only cleaves the peptide from the resin but alsocleaves all remaining side chain protecting groups and also the α-aminoprotecting group at the N-terminus if it was not previously removed toobtain the peptide in the form of the free acid. If Met is present inthe sequence, the Boc protecting group is preferably first removed usingtrifluoroacetic acid (TFA)/ethanedithiol prior to cleaving the peptidefrom the resin with HF to eliminate potential S-alkylation. When usinghydrogen fluoride or TFA for cleaving, one or more scavengers such asanisole, cresol, dimethyl sulfide and methylethyl sulfide are includedin the reaction vessel.

Cyclization of the linear peptide is preferably affected, as opposed tocyclizing the peptide while a part of the peptido-resin, to create bondsbetween Cys residues. To effect such a disulfide cyclizing linkage,fully protected peptide can be cleaved from a hydroxymethylated resin ora chloromethylated resin support by ammonolysis, as is well known in theart, to yield the fully protected amide intermediate, which isthereafter suitably cyclized and deprotected. Alternatively,deprotection, as well as cleavage of the peptide from the above resinsor a benzhydrylamine (BHA) resin or a methylbenzhydrylamine (MBHA), cantake place at 0° C. with hydrofluoric acid (HF) or TFA, followed byoxidation as described above.

The peptides are also synthesized using an automatic synthesizer. Aminoacids are sequentially coupled to an MBHA Rink resin (typically 100 mgof resin) beginning at the C-terminus using an Advanced Chemtech 357Automatic Peptide Synthesizer. Couplings are carried out using1,3-diisopropylcarbodimide in N-methylpyrrolidinone (NMP) or by 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate(HBTU) and diethylisopro-pylethylamine (DIEA). The FMOC protecting groupis removed by treatment with a 20% solution of piperidine indimethylformamide(DMF). Resins are subsequently washed with DMF (twice),followed by methanol and NMP.

Muteins, analogs or active fragments, of the foregoing conotoxinpeptides are also contemplated here. See, e.g., Hammerland et al, Eur.J. Pharmacol., 226, pp. 239-244 (1992). Derivative muteins, analogs oractive fragments of the conotoxin peptides may be synthesized accordingto known techniques, including conservative amino acid substitutions,such as outlined in U.S. Pat. Nos. 5,545,723 (see particularly col. 2,line 50—col. 3, line 8); 5,534,615 (see particularly col. 19, line45—col. 22, line 33); and 5,364,769 (see particularly col. 4, line55—col. 7, line 26), each herein incorporated by reference.

Pharmaceutical compositions containing a compound of the presentinvention or its pharmaceutically acceptable salts as the activeingredient can be prepared according to conventional pharmaceuticalcompounding techniques. See, for example, Remington's PharmaceuticalSciences, 18th Ed. (1990, Mack Publishing Co., Easton, Pa.). Typically,an antagonistic amount of the active ingredient will be admixed with apharmaceutically acceptable carrier. The carrier may take a wide varietyof forms depending on the form of preparation desired foradministration, e.g., intravenous, oral or parenteral. The compositionsmay further contain antioxidizing agents, stabilizing agents,preservatives and the like. For examples of delivery methods see U.S.Pat. No. 5,844,077, incorporated herein by reference.

For oral administration, the compounds can be formulated into solid orliquid preparations such as capsules, pills, tablets, lozenges, melts,powders, suspensions or emulsions. In preparing the compositions in oraldosage form, any of the usual pharmaceutical media may be employed, suchas, for example, water, glycols, oils, alcohols, flavoring agents,preservatives, coloring agents, suspending agents, and the like in thecase of oral liquid preparations (such as, for example, suspensions,elixirs and solutions); or carriers such as starches, sugars, diluents,granulating agents, lubricants, binders, disintegrating agents and thelike in the case of oral solid preparations (such as, for example,powders, capsules and tablets). Because of their ease in administration,tablets and capsules represent the most advantageous oral dosage unitform, in which case solid pharmaceutical carriers are obviouslyemployed. If desired, tablets may be sugar-coated or enteric-coated bystandard techniques. The active agent can be encapsulated to make itstable to passage through the gastrointestinal tract while at the sametime allowing for passage across the blood brain barrier. See forexample, WO 96/11698.

For parenteral administration, the compound may be dissolved in apharmaceutical carrier and administered as either a solution or asuspension. Illustrative of suitable carriers are water, saline,dextrose solutions, fructose solutions, ethanol, or oils of animal,vegetative or synthetic origin. The carrier may also contain otheringredients, for example, preservatives, suspending agents, solubilizingagents, buffers and the like. When the compounds are being administeredintrathecally, they may also be dissolved in cerebrospinal fluid.

The active agent is preferably administered in an therapeuticallyeffective amount. The actual amount administered, and the rate andtime-course of administration, will depend on the nature and severity ofthe condition being treated. Prescription of treatment, e.g. decisionson dosage, timing, etc., is within the responsibility of generalpractitioners or spealists, and typically takes account of the disorderto be treated, the condition of the individual patient, the site ofdelivery, the method of administration and other factors known topractitioners. Examples of techniques and protocols can be found inRemington's Pharmaceutical Sciences. Typically the active agents of thepresent invention exhibit their effect at a dosage range from about0.001 mg/kg to about 250 mg/kg, preferably from about 0.01 mg/kg toabout 100 mg/kg of the active ingredient, more preferably from a bout0.05 mg/kg to about 75 mg/kg. A suitable dose can be administered inmultiple sub-doses per day. Typically, a dose or sub-dose may containfrom about 0.1 mg to about 500 mg of the active ingredient per unitdosage form. A more preferred dosage will contain from about 0.5 mg toabout 100 mg of active ingredient per unit dosage form. Dosages aregenerally initiated at lower levels and increased until desired effectsare achieved.

Alternatively, targeting therapies may be used to deliver the activeagent more specifically to certain types of cell, by the use oftargeting systems such as antibodies or cell specific ligands. Targetingmay be desirable for a variety of reasons, e.g. if the agent isunacceptably toxic, or if it would otherwise require too high a dosage,or if it would not otherwise be able to enter the target cells.

The active agents, which are peptides, can also be administered in acell based delivery system in which a DNA sequence encoding an activeagent is introduced into cells designed for implantation in the body ofthe patient, especially in the spinal cord region. Suitable deliverysystems are described in U.S. Pat. No. 5,550,050 and published PCTApplication Nos. WO 92/19195, WO 94/25503, WO 95/01203, WO 95/05452, WO96/02286, WO 96/02646, WO 96/40871, WO 96/40959 and WO 97/12635.Suitable DNA sequences can be prepared synthetically for each activeagent on the basis of the developed sequences and the known geneticcode.

EXAMPLES

The present invention is described by reference to the followingExamples, which are offered by way of illustration and are not intendedto limit the invention in any manner. Standard techniques well known inthe art or the techniques specifically described below were utilized.

Example 1 Isolation of τ-Conotoxins

Crude venom was extracted from venom ducts (Cruz et al., 1976), and thecomponents were purified as previously described (Cartier et al., 1996).The crude extract from venom ducts was purified by reverse phase liquidchromatography (RPLC) using a Vydac Cl₈ semi-preparative column (10×250mm). Further purification of bioactive peaks was done on a Vydac C₁₈analytical column (4.6×220 mm). The effluents were monitored at 220 nm.Peaks were collected, and aliquots were assayed for activity.

The amino acid sequence of the purified peptides were determined bystandard methods. The purified peptides were reduced and alkylated priorto sequencing by automated Edman degradation on an Applied Biosystems477A Protein Sequencer with a 120A Analyzer (DNA/Peptide Facility,University of Utah) (Martinez et al., 1995; Shon et al., 1994).

In accordance with this method, peptides AuVA, AuVB and PVA wereobtained.

Example 2 Synthesis of conopeptides

The synthesis of conopeptides, either the mature toxins or the precursorpeptides, was separately performed using conventional protectionchemistry as described by Cartier et al. (1996). Briefly, the linearchains were built on Rink amide resin by Fmoc procedures with2-(1H-benzotriol-1-yl)-1,1,3,3,-tetramethyluronium tetrafluoroboratedcoupling using an ABI model 430A peptide synthesizer with amino acidderivatives purchased from Bachem (Torrence, Calif.). Orthogonalprotection was used on cysteines: two cysteines were protected as thestable Cys(S-acetamidomethyl), while the other two cysteines wereprotected as the acid-labile Cys(S-trityl). After removal of theterminal Fmoc protecting group and cleavage of the peptides from theresins, the released peptides were precipitated by filtering thereaction mixture into −10° C. methyl t-butyl ether, which removed theprotecting groups except the Cys(S-acetamidomethyl). The peptides weredissolved in 0.1% TFA and 60% acetonitrile and purified by RPLC on aVydac C₁₈ preparative column (22×250 mm) and eluted at a flow rate of 20mL/min with a gradient of acetonitrile in 0.1% TFA.

The disulfide bridges in the three conopeptides were formed as describedin Cartier et al. (1996). Briefly, the disulfide bridges between onepair of cysteines were formed by air oxidation which was judged to becomplete by analytical RPLC. The monocyclic peptides were purified byRPLC on a Vydac C₁₈ preparative column (22×250 mm) and eluted with agradient of acetonitrile in 0.1% TFA. Removal of S-acetamidomethylgroups and formation of the disulfide bridge between the other pair ofcysteines were carried out simultaneously be iodine oxidation. Thecyclic peptides were purified by RPLC on a Vydac C₁₈ preparative column(22×250 mm) and eluted with a gradient of acetonitrile in 0.1% TFA.

Example 3

Isolation of DNA Encoding τ-Conotoxins

DNA coding for τ-conotoxins was isolated and cloned in accordance withconventional techniques using general procedures well known in the art,such as described in Olivera et al. (1996). Alternatively, cDNAlibraries was prepared from Conus venom duct using conventionaltechniques. DNA from single clones was amplified by conventionaltechniques using primers which correspond approximately to the M13universal priming site and the M13 reverse universal priming site.Clones having a size of approximately 300-500 nucleotides were sequencedand screened for similarity in sequence to known r-conotoxins isolatedin Example 1. The DNA sequences and encoded propeptide sequences are setforth in Tables 1-15. DNA sequences coding for the mature toxin can alsobe prepared on the basis of the DNA sequences set forth in these Tables.

TABLE 1 DNA Sequence (SEQ ID NO:20) and Protein Sequence (SEQ ID NO:21)of Tx5.1 ggtactcaac gaacttcaag acacattctt ttcacctgga cacgggaagctgactacaag caga atg tgc tgt ctc cca gtg ttc gtc att ctt ctg ctg ctg attgca      Met Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Ala tctgca cct agc gtt gat gcc caa ccg aag acc aaa gat gat gtg ccc Ser Ala ProSer Val Asp Ala Gln Pro Lys Thr Lys Asp Asp Val Pro ctg gca cct ttg cacgat aat gca aag agt gca cta caa cat ttg aac Leu Ala Pro Leu His Asp AsnAla Lys Ser Ala Leu Gln His Leu Asn caa cgc tgc tgc caa aca ttc tat tggtgc tgt gtt caa ggg aaa Gln Arg Cys Cys Gln Thr Phe Tyr Trp Cys Cys ValGln Gly Lys tgaatttgga tgagacccct gcgaactgtc catggatgtg agatttggaaagcagactgt tcctttcgca cgtgttcgtg gaattttgaa tggtcgttaa caacacgctgccacttgcaa gctactatct ctctgtcctt tcatctgtgg aactggatga cctaacaactgaaatatcat agaaattttt cagtgggtat acactatgac catgtagtca gtaattacatcatttggacc ttttgaaata tttttcaaaa tgttaagatt tttcccccng gaaaggncttttgaagtaaa tatt

TABLE 2 DNA Sequence (SEQ ID NO:22) and Protein Sequence (SEQ ID NO:23)of G5.1 atg tgc tgt ctc cca gtc ttc gtc att ctt ctg ttg ctg att aca tctMet Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Thr Ser gca cctagc gtt gat gct cta ccg aag acc agg gat gat gtg ccc cta Ala Pro Ser ValAsp Ala Leu Pro Lys Thr Arg Asp Asp Val Pro Leu gca tct ttc cac ggt ggatat aat gca agg aga atc cta caa agg cgt Ala Ser Phe His Gly Gly Tyr AsnAla Arg Arg Ile Leu Gln Arg Arg cag ggc tgg tgc tgc aaa gaa aat att gcgtgc tgt ata tagtggtaac Gln Gly Trp Cys Cys Lys Glu Asn Ile Ala Cys CysIle gggaaatgac tttggatgag acccctgcaa actgtccctg gatgtgaaat ttggaaagtagactgttcct ttcgcgcgtg ttcgtggaat ttcaaatggt cgtcaacaac acactgctacttgcaaagct actatctctc tgtcctttca tctgtggaac tgggtgatct aacagctgaaatgtcgcaga aatttttcaa ttggtctata ctatgaccat gta

TABLE 3 DNA Sequence (SEQ ID NO:24) and Protein Sequence (SEQ ID NO:25)of Qc5.1 atg cgc tgt gtc cca gtc ttc atc att ctt ctg ctg ctg agt cca tctMet Arg Cys Val Pro Val Phe Ile Ile Leu Leu Leu Leu Ser Pro Ser gca cctagc gtt gat gcc cat ccg atg acc aaa gat gat gtg ccc cag Ala Pro Ser ValAsp Ala His Pro Met Thr Lys Asp Asp Val Pro Gln gca tca ttc cat gat gatgca aag cga acc cta caa gta cct tgg atg Ala Ser Phe His Asp Asp Ala LysArg Thr Leu Gln Val Pro Trp Met aaa cgc ggg tgc tgc gca agg ttg act tgctgc gtt gga cga Lys Arg Gly Cys Cys Ala Arg Leu Thr Cys Cys Val Gly Argtaaagggaaa tgactttgga tgagacccct gcgaactgtc cctggatgtg aaatttggacagcagactgc tcctttcgca cgtgttcgtg gaattttgaa tggtcgttaa caacacgctgccacttgcaa gctattatct ctctgtccct ttatctgtgg aactggataa tctaacaactgaaatgtcat tgaaaatttt caatggatat atattatgat ccatata

TABLE 4 DNA Sequence (SEQ ID NO:26) and Protein Sequence (SEQ ID NO:27)of Im5.1 aattcggaag ctgactacaa gcaga atg tac tgt ctc cca gtc ttc atc att                            Met Tyr Cys Leu Pro Val Phe Ile Ile ctt ctgctg ctg att tca tct gca cct agc act cct ccc caa cca agg Leu Leu Leu LeuIle Ser Ser Ala Pro Ser Thr Pro Pro Gln Pro Arg aac aaa gat cgt gtg cacctg ata tct tta ctc gat aat cac aag caa Asn Lys Asp Arg Val His Leu IleSer Leu Leu Asp Asn His Lys Gln atc cta caa aga gat tgg aac agt tgc tgtggg aaa aat cct ggt tgc Ile Leu Gln Arg Asp Trp Asn Ser Cys Cys Gly LysAsn Pro Gly Cys tgt cct tgg gga aaa tgactttgga tgagacccct gcaaactgtccctggatgtg Cys Pro Trp Gly Lys agatttggaa agcagaccgt ttgtggaattttgaatggtc gttaacaaca cgctgccact tgcaagctac aatctctctg tcctttcatctttggaactg gatgatcaaa caactgaaat gtcatagaaa tttttcaatg ggtatacaatatgtgggcat ttagtcagta attacatcat ttgg

TABLE 5 DNA Sequence (SEQ ID NO:28) and Protein Sequence (SEQ ID NO:29)of G5.2 atg tgc tgt ctc cca gtc ttc gtc att ctt ctg ttg ctg att aca tctMet Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Thr Ser gca cctagc gtt gat gct cta ccg aag acc agg gat gat gtg ccc cta Ala Pro Ser ValAsp Ala Leu Pro Lys Thr Arg Asp Asp Val Pro Leu gca tct ttc cac ggt ggatat aat gca agg aga atc cta caa agg cgt Ala Ser Phe His Gly Gly Tyr AsnAla Arg Arg Ile Leu Gln Arg Arg cag ggc tgg tgc tgc aaa gaa aat att gcgtgc tgt gta tagtggtaac Gln Gly Trp Cys Cys Lys Glu Asn Ile Ala Cys CysVal gggaaatgac tttggatgag acccctgcaa actgtccctg gatgtgaaat ttggaaagtagactgttcct ttcgcgcgtg ttcgtggaat ttcaaatggt cgtcaacaac acactgctacttgcaaagct actatctctc tgtcctttca tctgtggaac tgggtgatct aacagctgaaatgtcgcaga aatttttcaa ttggtctata ctatgaccat gtagtcag

TABLE 6 DNA Sequence (SEQ ID NO:30) and Protein Sequence (SEQ ID NO:31)of Tx5.2a atg cgc tgt ttc cca gtc ttc atc att ctt ctg ctg cta att gcatct Met Arg Cys Phe Pro Val Phe Ile Ile Leu Leu Leu Leu Ile Ala Ser gcacct tgc ttt gat gcc cga acg aag acc gat gat gat gtg ccc ctg Ala Pro CysPhe Asp Ala Arg Thr Lys Thr Asp Asp Asp Val Pro Leu tca tct ctc cgc gataat cta aag cga acg ata cga aca cgc ctg aac Ser Ser Leu Arg Asp Asn LeuLys Arg Thr Ile Arg Thr Arg Leu Asn ata cgc gag tgc tgc gag gat gga tggtgc tgt act gct gca ccc tta Ile Arg Glu Cys Cys Glu Asp Gly Trp Cys CysThr Ala Ala Pro Leu aca ggt cgt tagggataaa ggaaaatggc tttggatgagacccctgcga Thr Gly Arg attgtccctg gatgtgagat ttggaaagca gactgttcctttcgcacgtg ttcgtggaat ttcgaatggt cgttaacaac acgctgccac ttgcaagccaccatctctct gtcctttcgt atgtggaact gtatgatcta acaactgaaa tgtcagaaagttttcagtgg gtatacacta tgatcgtata

TABLE 7 DNA Sequence (SEQ ID NO:32) and Protein Sequence (SEQ ID NO:33)of Tx5.2b atg cgc tgt ttc cca gtc ttc atc att ctt ctg ttg cta att gcatct Met Arg Cys Phe Pro Val Phe Ile Ile Leu Leu Leu Leu Ile Ala Ser gcacct tgc ttt gat gcc cga acg aag acc gat gat gat gtg ccc ctg Ala Pro CysPhe Asp Ala Arg Thr Lys Thr Asp Asp Asp Val Pro Leu tca tct ctc cgc gataat cta aag cga acg ata cga aca cgc ctg aac Ser Ser Leu Arg Asp Asn LeuLys Arg Thr Ile Arg Thr Arg Leu Asn ata cgc ggg tgc tgc gag gat gga tggtgc tgt act gct gca ccc tta Ile Arg Gly Cys Cys Glu Asp Gly Trp Cys CysThr Ala Ala Pro Leu aca ggt cgt tagggataaa ggaaaatggc tttggatgagacccctgcaa Thr Gly Arg attgtccctg gatgtgagat ttggaaagca gactgttcctttcgcacgtg ttcgtggaat ttcgaatggt cgttaacaac acgctgccac ttgcaagccaccatctctct gtcctttcgt atgtggaact gtatgatcta acaactgaaa tgtcagaaagttttcagtgg gtatacacta tgatcgtata gtcagtaatt

TABLE 8 DNA Sequence (SEQ ID NO:34) and Protein Sequence (SEQ ID NO:35)of Mr5.1 atg cgc tgc ctc cca gtc ttc gtc att ctt ctg ctg ctg att gca tctMet Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Ala Ser gca cctagc gtt gat gcc cga ccg aag acc aaa gat gat atg ccc ctg Ala Pro Ser ValAsp Ala Arg Pro Lys Thr Lys Asp Asp Met Pro Leu gca tct ttc cat gat aatgca aag cga atc ctg caa ata ctt cag gac Ala Ser Phe His Asp Asn Ala LysArg Ile Leu Gln Ile Leu Gln Asp aga aat ggt tgc tgc aga gca gga gac tgctgt tca cga ttt gag ata Arg Asn Gly Cys Cys Arg Ala Gly Asp Cys Cys SerArg Phe Glu Ile aag gaa aat gac ttt gga tgagacccct gcaaactgtc cttggatgtgLys Glu Asn Asp Phe Gly agatttggaa agcagactgt tcctttcgca cgtgttcgtggaatttcgaa tggtcgttaa caacacgctg ccacttgcaa gctactatct ctctgtccttttgtctgtgg aactgtatga tcaaacaact gaaatgtcat agaaattttt cagtgggtaaacactatgac catgta

TABLE 9 DNA Sequence (SEQ ID NO:36) and Protein Sequence (SEQ ID NO:37)of Mr5.2 ga atg cgc tgc ctc cca gtc ttc gtc att ctt ctg ctg ctg att gca   Met Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Ala tct gcacct agc gtt gat gcc cga ccg aag acc aaa gat gat atg ccc Ser Ala Pro SerVal Asp Ala Arg Pro Lys Thr Lys Asp Asp Met Pro ctg gca tct ttc cac gataat gca aag cga atc ctg caa ata ctt cag Leu Ala Ser Phe His Asp Asn AlaLys Arg Ile Leu Gln Ile Leu Gln gac aga aat gct tgc tgc ata gta agg cagtgc tgt tgatgatttg Asp Arg Asn Ala Cys Cys Ile Val Arg Gln Cys Cysagataaagga aaatgacttt ggatgagacc cctgcaaact gtccctggat gtgagatttggaaagcagac tgttcctttc gcacgtgttc gtggaatttc gaatggtcgt taacaacacgctgccacttg caagctacta tctctctgtc ctttcatctg tggaactgta tgatcaaacaactgaaatgt catagaaatt tttcagtggg taaacactat gatcatgtag tcagtaattacatcatttgg aattccatca agcttatcga taccgtcgac ctcgaggggg ggcccggt

TABLE 10 DNA Sequence (SEQ ID NO:38) and Protein Sequence (SEQ ID NO:39)of Mr5.3 atg cgc tgc ctc cca gtc ttt gtc att ctt ctg ctg ctg att gca tctMet Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Ala Ser gca cctagc gtt gat gcc cga ccg aag acc aaa gat gat atg ccc ctg Ala Pro Ser ValAsp Ala Arg Pro Lys Thr Lys Asp Asp Met Pro Leu gca tct ttc cat gat aatgca aag cga atc ctg caa ata ctt cag gac Ala Ser Phe His Asp Asn Ala LysArg Ile Leu Gln Ile Leu Gln Asp aga aat ggt tgc tgc aga gca gga gac tgctgt tca tgatttgaga Arg Asn Gly Cys Cys Arg Ala Gly Asp Cys Cys Sertaaagggaaa tgactttgga tgagacccct gcaaactgtc cttggatgtg agatttggaaagcagactgt tcctttcgca cgtgttcgtg gaatttcgaa tggtcgttaa caacacgctgccacttgcaa gctactatct ctctgtcctt tcatctgtgg aactgtatga tcaaacaact

TABLE 11 DNA Sequence (SEQ ID NO:40) and Protein Sequence (SEQ ID NO:41)of Ca5.1 atg cgc tgt ctc ccg gtc ttc atc att ctt ctg ctg ctg att gca tctMet Arg Cys Leu Pro Val Phe Ile Ile Leu Leu Leu Leu Ile Ala Ser gca cctggc gtt gat gcc caa ccg aag acc aaa tat aat gcg ccc ctg Ala Pro Gly ValAsp Ala Gln Pro Lys Thr Lys Tyr Asn Ala Pro Leu aca tct ctc cac gat aatgca aag ggt ata cta caa gaa cat tgg aac Thr Ser Leu His Asp Asn Ala LysGly Ile Leu Gln Glu His Trp Asn aaa cgc tgc tgc ccc aga agg ctt gcc tgctgt att ata gga cgg aaa Lys Arg Cys Cys Pro Arg Arg Leu Ala Cys Cys IleIle Gly Arg Lys tggatgattt tgggtgagat ccctgcaaac tgtccctgga tttgaattttggaaagcaga ctgttccttt cgcacgtgtt cgtggaattt cgaatggtcg ttaacaacacgctgccactt gcaagctact atctctctgt cctttttctc tgtgaaactg gatggtctaacaactgaaat gtcatagaaa attttcaatg ggtatactct atgaccatct a

TABLE 12 DNA Sequence (SEQ ID NO:42) and Protein Sequence (SEQ ID NO:43)of Ca5.2 atg cgc tgt ctc cca gtc ttc atc att ctt ctg ctg ctg att gca tctMet Arg Cys Leu Pro Val Phe Ile Ile Leu Leu Leu Leu Ile Ala Ser gca cctggc gtt gat gcc caa ccg aag acc aaa tat gat gcg ccc ctg Ala Pro Gly ValAsp Ala Gln Pro Lys Thr Lys Tyr Asp Ala Pro Leu aca tct ctc cac gat aatgca aag ggt ata cta caa gaa cat tgg aac Thr Ser Leu His Asp Asn Ala LysGly Ile Leu Gln Glu His Trp Asn aaa cgc tgc tgc ccc aac aag cct tgc tgtttt ata gga agg aaa Lys Arg Cys Cys Pro Asn Lys Pro Cys Cys Phe Ile GlyArg Lys tgaatgattt tgggtgagac ccctgcaaac tgtccctgga tttgaattttggaaagcaga ctgttccttt cgcacgtgtt cgtggaattt cgaatggtcg ttaacaacacgctgccactt gcaagctact atctctctgt cctttttctc tgtgaaactg gatggtctaacaactgagat gtcatagaaa attttcaatc ggtgtactct atgaccatct a

TABLE 13 DNA Sequence (SEQ ID NO:44) and Protein Sequence (SEQ ID NO:45)of Qc5.2 atg cgc tgt gtc cca gtc ttc atc att ctt ctg ctg ctg agt cca tctMet Arg Cys Val Pro Val Phe Ile Ile Leu Leu Leu Leu Ser Pro Ser gca cctagc gtt gat gcc cat ccg atg acc aaa gat gat gta ccc cag Ala Pro Ser ValAsp Ala His Pro Met Thr Lys Asp Asp Val Pro Gln gca tct ctc cat gat gatgca aag cga acc cta caa gta cct tgg atg Ala Ser Leu His Asp Asp Ala LysArg Thr Leu Gln Val Pro Trp Met aaa cgc ggg tgc tgc gca atg ttg act tgctgc gtt gga cga Lys Arg Gly Cys Cys Ala Met Leu Thr Cys Cys Val Gly Argtaaagggaaa tgactttgga tgagacccct acgaactgtc cctggatgtg aaatttggacagcagactgc tcctttcgca cgtgttcgtg gaatttcgaa tggtcgttaa caacacgctgccacttgcaa gctattatct ctctgtccct ttatctgtgg aactggataa tctaacaactgaaacgtcat tgaaaatttt caatggatat atattatgat ccatata

TABLE 14 DNA Sequence (SEQ ID NO:46) and Protein Sequence (SEQ ID NO:47)of Gm5.1 gggcaggtac tcaacgaact tcaggacaca ttcttttcac ctggacacgggaaactgact ataagcaga atg cgc tac cta cca gtc ttc gtc att ctt ctg ctg ctgatt           Met Arg Tyr Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ilegca tct ata cct agc gat act gtc caa ctg aag acc aaa gat gat atg Ala SerIle Pro Ser Asp Thr Val Gln Leu Lys Thr Lys Asp Asp Met ccc ctg gca tctttc cac ggt aat gga aga cga atc ctg cga atg ctt Pro Leu Ala Ser Phe HisGly Asn Gly Arg Arg Ile Leu Arg Met Leu tca aac aaa cgc tta tgc tgt gtcacc gag gat tgg tgc tgt gaa tgg Ser Asn Lys Arg Leu Cys Cys Val Thr GluAsp Trp Cys Cys Glu Trp tgg taaaggaaaa tgactttgga tgagacccct gcaaactgtttctggatgtg Trp agatttggaa agcagactgt tctttcgcac gtattcgtga aatttcgaatggtcgttaac aacacgctgc cacttgcaag ctgctatctc tctgtctttt catctgtggaactgtatgat ctaacaactg aaatgtcata gacatttttc attgggtata cactatgaccatgtagccag taattacatc atttggacct tttggatatt tttcagtatg taagtgtgttcccttaaaaa gtcctttgta attatgtatt ttaanaattt angttttgca cataaattgtaaaacgctgt cctttctgtt gntcctacat cantggtggg gaaaagnaaa atgtttggccntggtcaaat ttaaataatn accctgccgt ttnaatgcng ttattantgg tattttnaacnttgnacggt taaactt

TABLE 15 DNA Sequence (SEQ ID NO:48) and Protein Sequence (SEQ ID NO:49)of Gm5.2 ga atg cgc tgt ctc cca gtc ttc gtc att ctt ctg ctg ctg att gca   Met Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Ala tct gcacct agc gtt gat gcc caa ccg aag acc aaa gat gat gtg ccc Ser Ala Pro SerVal Asp Ala Gln Pro Lys Thr Lys Asp Asp Val Pro ctg gca cct ttg cac gataat ata agg agt act cta caa aca ctt cgg Leu Ala Pro Leu His Asp Asn IleArg Ser Thr Leu Gln Thr Leu Arg aag aaa gtc tgc tgc cgc cca gtg cag gattgc tgt tca ggg aaa Lys Lys Val Cys Cys Arg Pro Val Gln Asp Cys Cys SerGly Lys tgaagggaaa tgaatttgga tgagacccct gcgaactgtc cctggatgtgagatttggaa agcagactgt tcctttcgca cgtgttcgtg gaatttcgaa tggtcgttaacaacacgctg ccacttgcaa gctactatct ctctgtcctt tcatctgcgg aactggatgacctaaagctt gtgatc

Example 4 Biological Activity of τ-Conotoxins

The biological activity of τ-conotoxin peptides at the acetylcholinereceptor was tested in the fluorescence assay as described byCornell-Bell et al. (1999). Briefly, primary cortical cells are exposedto acetylcholine in the presence or absence of a τ-conotoxin peptide.Acetylcholine causes the primary cortical cells to flux calcium, whichis measured by increases in fluorescence in cells loaded with Fluo-3, acalcium imaging dye. The τ-conotoxin peptide AuVA inhibited the responseof primary cortical cells to acetylcholine at low concentrations (10 pM)at 15 seconds following exposure to the peptide and acetylcholine. Thisstudy shows that the τ-conotoxin peptide acts at the acetylcholinereceptor.

Example 5 Effect of τ-Conotoxins in a Pain Model

The effect of τ-conotoxin peptides for use in treating pain was bytesting in two pain models, the first being the hind-paw licking model(Woolfe and MacDonald, 1944; Plummer et al., 1991; Suh et al., 1992;Plone et al., 1996) and the second being the accelerating roto-rodmodel. In the hind-paw licking model, it was found that 10 nmol ofτ-conotoxin peptide AuVA increased the latency to initiate hind-pawlicking in mice on a 55° C. hot plate 15 minutes following freehandintracerebroventricular (i.c.v.) injection. It was further found that 1nmol τ-conotoxin peptide AuVA did not have any effect in this model. Inthe accelerating roto-rod model, it was found that τ-conotoxin peptideAuVA produced impairment of motor performance following injection ofτ-conotoxin peptide AuVA. The effects seen in these models demonstratesthat the τ-conotoxin peptides have analgesic properties.

It will be appreciated that the methods and compositions of the instantinvention can be incorporated in the form of a variety of embodiments,only a few of which are disclosed herein. It will be apparent to theartisan that other embodiments exist and do not depart from the spiritof the invention. Thus, the described embodiments are illustrative andshould not be construed as restrictive.

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49 1 23 PRT Artificial Sequence Description of ArtificialSequenceGeneric Sequence for Tau Conopeptides 1 Xaa Xaa Xaa Xaa Cys CysXaa Xaa Xaa Xaa Xaa Cys Cys Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa XaaXaa Xaa 20 2 11 PRT Conus aulicus PEPTIDE (4)..(8) Xaa at residue 4 isPro or hydroxy-Pro; Xaa at residue 8 is Tyr, nor-Tyr, mono-halo-Tyr,di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr or nitro-Tyr. 2 Phe Cys Cys XaaVal Ile Arg Xaa Cys Cys Xaa 1 5 10 3 11 PRT Conus aulicus PEPTIDE(4)..(8) Xaa at residue 4 is Pro or hydroxy-Pro; Xaa at residue 8 isTyr, nor-Tyr, mono-halo-Tyr, di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr ornitro-Tyr. 3 Phe Cys Cys Xaa Phe Ile Arg Xaa Cys Cys Xaa 1 5 10 4 10 PRTConus textile PEPTIDE (6)..(7) Xaa at residue 6 is Tyr, mono-halo-Tyr,di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr or nitro-Tyr; Xaa at residue 7is Trp (D or L), neo-Trp or halo-Trp (D or L). 4 Cys Cys Gln Thr Phe XaaXaa Cys Cys Gln 1 5 10 5 13 PRT Conus geographus PEPTIDE (1)..(3) Xaa atresidue 1 is Gln or pyro-Glu; Xaa at residue 3 is Trp (D or L), neo-Trpor halo-Trp (D or L); Xaa at residue 6 is Lys, N-methyl-Lys,N,N-dimethyl-Lys or N,N,N-trimethyl-Lys. 5 Xaa Gly Xaa Cys Cys Xaa XaaAsn Ile Ala Cys Cys Ile 1 5 10 6 10 PRT Conus quercinus 6 Gly Cys CysAla Arg Leu Thr Cys Cys Val 1 5 10 7 12 PRT Conus purpurascens PEPTIDE(5)..(6) Xaa at residue 5 is Pro or hydroxy-Pro; Xaa at residue 6 isLys, N-methyl-Lys, N,N-dimethyly-Lys or N,N,N-trimethyl-Lys. 7 Asn GlyCys Cys Xaa Lys Gln Met Arg Cys Cys Thr 1 5 10 8 15 PRT Conus imperialisPEPTIDE (2)..(15) Xaa at residues 2 and 15 is Trp (D or L), neo-Trp orhalo-Trp (D or L); Xaa at residue 8 is Lys, N-methyl-Lys,N,N-dimethyl-Lys or N,N,N-trimethyl-Lys; Xaa at residues 10 and 14 isPro or hydroxy-Pro. 8 Asp Xaa Asn Ser Cys Cys Gly Xaa Asn Xaa Gly CysCys Xaa Xaa 1 5 10 15 9 13 PRT Conus geographus PEPTIDE (1)..(6) Xaa atresidue 1 is Gln or pyro-Glu; Xaa at residue 2 is Trp (D or L), neo-Trpor halo-Trp (D or L); Xaa at residue 6 is Lys, N-methyl-Lys,N,N,-dimethyl-Lys or N,N,N-trimethyl-Lys. 9 Xaa Gly Xaa Cys Cys Xaa XaaAsn Ile Arg Cys Cys Val 1 5 10 10 15 PRT Conus textile PEPTIDE (1)..(13)Xaa at residues 1 and 4 is Glu or gamma-carboxy-Glu; Xaa at residue 7 isTrp (D or L), neo-Trp or halo-Trp (D or L); Xaa at residue 13 is Pro orhydroxy-Pro. 10 Xaa Cys Cys Xaa Asp Gly Xaa Cys Cys Thr Ala Ala Xaa LeuThr 1 5 10 15 11 15 PRT Conus textile PEPTIDE (4)..(13) ; Xaa at residue4 is Glu or gamma-carboxy-Glu; Xaa at residue 7 is Trp (D or L) neo-Trpor halo-Trp (D or L); Xaa at residue 13 is Pro or hydroxy-Pro. 11 GlyCys Cys Xaa Asp Gly Xaa Cys Cys Thr Ala Ala Xaa Leu Thr 1 5 10 15 12 20PRT Conus marmoreus PEPTIDE (14)..(17) Xaa at residue 14 and 17 is Gluor gamma-carboxy-Glu; Xaa at residue 16 is Lys, N-methyl-Lys,N,N-dimethyl-Lys or N,N,N-trimethyl-Lys. 12 Asn Gly Cys Cys Arg Ala GlyAsp Cys Cys Ser Arg Phe Xaa Ile Xaa 1 5 10 15 Xaa Asn Asp Phe 20 13 10PRT Conus marmoreus 13 Asn Ala Cys Cys Ile Val Arg Gln Cys Cys 1 5 10 1411 PRT Conus marmoreus 14 Asn Gly Cys Cys Arg Ala Gly Asp Cys Cys Ser 15 10 15 11 PRT Conus caracteristicus PEPTIDE (3) Xaa at residue 3 is Proor hydroxy-Pro. 15 Cys Cys Xaa Arg Arg Leu Ala Cys Cys Ile Ile 1 5 10 1610 PRT Conus caracteristicus PEPTIDE (3)..(6) Xaa at residue 3 and 6 isPro or hydroxy-Pro; Xaa at residue 5 is Lys, N-methyl-Lys,N,N-dimethyl-Lys or N,N,N-trimethyl-Lys. 16 Cys Cys Xaa Asn Xaa Xaa CysCys Phe Ile 1 5 10 17 10 PRT Conus quercinus 17 Gly Cys Cys Ala Met LeuThr Cys Cys Val 1 5 10 18 13 PRT Conus gloriamaris PEPTIDE (6)..(13) Xaaat residue 6 and 11 is Glu or gamma-carboxy-Glu; Xaa at residues 8, 12and 13 is Trp (D or L), neo-Trp or halo-Trp (D or L). 18 Leu Cys Cys ValThr Xaa Asp Xaa Cys Cys Xaa Xaa Xaa 1 5 10 19 11 PRT Conus gloriamarisPEPTIDE (5) Xaa at residue 5 is Pro or hydroxy-Pro. 19 Val Cys Cys ArgXaa Val Gln Asp Cys Cys Ser 1 5 10 20 554 DNA Conus textile CDS(65)..(250) 20 ggtactcaac gaacttcaag acacattctt ttcacctgga cacgggaagctgactacaag 60 caga atg tgc tgt ctc cca gtg ttc gtc att ctt ctg ctg ctgatt gca 109 Met Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Ala1 5 10 15 tct gca cct agc gtt gat gcc caa ccg aag acc aaa gat gat gtgccc 157 Ser Ala Pro Ser Val Asp Ala Gln Pro Lys Thr Lys Asp Asp Val Pro20 25 30 ctg gca cct ttg cac gat aat gca aag agt gca cta caa cat ttg aac205 Leu Ala Pro Leu His Asp Asn Ala Lys Ser Ala Leu Gln His Leu Asn 3540 45 caa cgc tgc tgc caa aca ttc tat tgg tgc tgt gtt caa ggg aaa 250Gln Arg Cys Cys Gln Thr Phe Tyr Trp Cys Cys Val Gln Gly Lys 50 55 60tgaatttgga tgagacccct gcgaactgtc catggatgtg agatttggaa agcagactgt 310tcctttcgca cgtgttcgtg gaattttgaa tggtcgttaa caacacgctg ccacttgcaa 370gctactatct ctctgtcctt tcatctgtgg aactggatga cctaacaact gaaatatcat 430agaaattttt cagtgggtat acactatgac catgtagtca gtaattacat catttggacc 490ttttgaaata tttttcaaaa tgttaagatt tttcccccng gaaaggnctt ttgaagtaaa 550tatt 554 21 62 PRT Conus textile 21 Met Cys Cys Leu Pro Val Phe Val IleLeu Leu Leu Leu Ile Ala Ser 1 5 10 15 Ala Pro Ser Val Asp Ala Gln ProLys Thr Lys Asp Asp Val Pro Leu 20 25 30 Ala Pro Leu His Asp Asn Ala LysSer Ala Leu Gln His Leu Asn Gln 35 40 45 Arg Cys Cys Gln Thr Phe Tyr TrpCys Cys Val Gln Gly Lys 50 55 60 22 416 DNA Conus geographus CDS(1)..(183) 22 atg tgc tgt ctc cca gtc ttc gtc att ctt ctg ttg ctg attaca tct 48 Met Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile ThrSer 1 5 10 15 gca cct agc gtt gat gct cta ccg aag acc agg gat gat gtgccc cta 96 Ala Pro Ser Val Asp Ala Leu Pro Lys Thr Arg Asp Asp Val ProLeu 20 25 30 gca tct ttc cac ggt gga tat aat gca agg aga atc cta caa aggcgt 144 Ala Ser Phe His Gly Gly Tyr Asn Ala Arg Arg Ile Leu Gln Arg Arg35 40 45 cag ggc tgg tgc tgc aaa gaa aat att gcg tgc tgt ata tagtggtaac193 Gln Gly Trp Cys Cys Lys Glu Asn Ile Ala Cys Cys Ile 50 55 60gggaaatgac tttggatgag acccctgcaa actgtccctg gatgtgaaat ttggaaagta 253gactgttcct ttcgcgcgtg ttcgtggaat ttcaaatggt cgtcaacaac acactgctac 313ttgcaaagct actatctctc tgtcctttca tctgtggaac tgggtgatct aacagctgaa 373atgtcgcaga aatttttcaa ttggtctata ctatgaccat gta 416 23 61 PRT Conusgeographus 23 Met Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu IleThr Ser 1 5 10 15 Ala Pro Ser Val Asp Ala Leu Pro Lys Thr Arg Asp AspVal Pro Leu 20 25 30 Ala Ser Phe His Gly Gly Tyr Asn Ala Arg Arg Ile LeuGln Arg Arg 35 40 45 Gln Gly Trp Cys Cys Lys Glu Asn Ile Ala Cys Cys Ile50 55 60 24 413 DNA Conus quercinus CDS (1)..(186) 24 atg cgc tgt gtccca gtc ttc atc att ctt ctg ctg ctg agt cca tct 48 Met Arg Cys Val ProVal Phe Ile Ile Leu Leu Leu Leu Ser Pro Ser 1 5 10 15 gca cct agc gttgat gcc cat ccg atg acc aaa gat gat gtg ccc cag 96 Ala Pro Ser Val AspAla His Pro Met Thr Lys Asp Asp Val Pro Gln 20 25 30 gca tca ttc cat gatgat gca aag cga acc cta caa gta cct tgg atg 144 Ala Ser Phe His Asp AspAla Lys Arg Thr Leu Gln Val Pro Trp Met 35 40 45 aaa cgc ggg tgc tgc gcaagg ttg act tgc tgc gtt gga cga 186 Lys Arg Gly Cys Cys Ala Arg Leu ThrCys Cys Val Gly Arg 50 55 60 taaagggaaa tgactttgga tgagacccct gcgaactgtccctggatgtg aaatttggac 246 agcagactgc tcctttcgca cgtgttcgtg gaattttgaatggtcgttaa caacacgctg 306 ccacttgcaa gctattatct ctctgtccct ttatctgtggaactggataa tctaacaact 366 gaaatgtcat tgaaaatttt caatggatat atattatgatccatata 413 25 62 PRT Conus quercinus 25 Met Arg Cys Val Pro Val Phe IleIle Leu Leu Leu Leu Ser Pro Ser 1 5 10 15 Ala Pro Ser Val Asp Ala HisPro Met Thr Lys Asp Asp Val Pro Gln 20 25 30 Ala Ser Phe His Asp Asp AlaLys Arg Thr Leu Gln Val Pro Trp Met 35 40 45 Lys Arg Gly Cys Cys Ala ArgLeu Thr Cys Cys Val Gly Arg 50 55 60 26 435 DNA Conus imperialis CDS(26)..(211) 26 aattcggaag ctgactacaa gcaga atg tac tgt ctc cca gtc ttcatc att 52 Met Tyr Cys Leu Pro Val Phe Ile Ile 1 5 ctt ctg ctg ctg atttca tct gca cct agc act cct ccc caa cca agg 100 Leu Leu Leu Leu Ile SerSer Ala Pro Ser Thr Pro Pro Gln Pro Arg 10 15 20 25 aac aaa gat cgt gtgcac ctg ata tct tta ctc gat aat cac aag caa 148 Asn Lys Asp Arg Val HisLeu Ile Ser Leu Leu Asp Asn His Lys Gln 30 35 40 atc cta caa aga gat tggaac agt tgc tgt ggg aaa aat cct ggt tgc 196 Ile Leu Gln Arg Asp Trp AsnSer Cys Cys Gly Lys Asn Pro Gly Cys 45 50 55 tgt cct tgg gga aaatgactttgga tgagacccct gcaaactgtc cctggatgtg 251 Cys Pro Trp Gly Lys 60agatttggaa agcagaccgt ttgtggaatt ttgaatggtc gttaacaaca cgctgccact 311tgcaagctac aatctctctg tcctttcatc tttggaactg gatgatcaaa caactgaaat 371gtcatagaaa tttttcaatg ggtatacaat atgtgggcat ttagtcagta attacatcat 431ttgg 435 27 62 PRT Conus imperialis 27 Met Tyr Cys Leu Pro Val Phe IleIle Leu Leu Leu Leu Ile Ser Ser 1 5 10 15 Ala Pro Ser Thr Pro Pro GlnPro Arg Asn Lys Asp Arg Val His Leu 20 25 30 Ile Ser Leu Leu Asp Asn HisLys Gln Ile Leu Gln Arg Asp Trp Asn 35 40 45 Ser Cys Cys Gly Lys Asn ProGly Cys Cys Pro Trp Gly Lys 50 55 60 28 421 DNA Conus geographus CDS(1)..(183) 28 atg tgc tgt ctc cca gtc ttc gtc att ctt ctg ttg ctg attaca tct 48 Met Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile ThrSer 1 5 10 15 gca cct agc gtt gat gct cta ccg aag acc agg gat gat gtgccc cta 96 Ala Pro Ser Val Asp Ala Leu Pro Lys Thr Arg Asp Asp Val ProLeu 20 25 30 gca tct ttc cac ggt gga tat aat gca agg aga atc cta caa aggcgt 144 Ala Ser Phe His Gly Gly Tyr Asn Ala Arg Arg Ile Leu Gln Arg Arg35 40 45 cag ggc tgg tgc tgc aaa gaa aat att gcg tgc tgt gta tagtggtaac193 Gln Gly Trp Cys Cys Lys Glu Asn Ile Ala Cys Cys Val 50 55 60gggaaatgac tttggatgag acccctgcaa actgtccctg gatgtgaaat ttggaaagta 253gactgttcct ttcgcgcgtg ttcgtggaat ttcaaatggt cgtcaacaac acactgctac 313ttgcaaagct actatctctc tgtcctttca tctgtggaac tgggtgatct aacagctgaa 373atgtcgcaga aatttttcaa ttggtctata ctatgaccat gtagtcag 421 29 61 PRT Conusgeographus 29 Met Cys Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu IleThr Ser 1 5 10 15 Ala Pro Ser Val Asp Ala Leu Pro Lys Thr Arg Asp AspVal Pro Leu 20 25 30 Ala Ser Phe His Gly Gly Tyr Asn Ala Arg Arg Ile LeuGln Arg Arg 35 40 45 Gln Gly Trp Cys Cys Lys Glu Asn Ile Ala Cys Cys Val50 55 60 30 431 DNA Conus textile CDS (1)..(201) 30 atg cgc tgt ttc ccagtc ttc atc att ctt ctg ctg cta att gca tct 48 Met Arg Cys Phe Pro ValPhe Ile Ile Leu Leu Leu Leu Ile Ala Ser 1 5 10 15 gca cct tgc ttt gatgcc cga acg aag acc gat gat gat gtg ccc ctg 96 Ala Pro Cys Phe Asp AlaArg Thr Lys Thr Asp Asp Asp Val Pro Leu 20 25 30 tca tct ctc cgc gat aatcta aag cga acg ata cga aca cgc ctg aac 144 Ser Ser Leu Arg Asp Asn LeuLys Arg Thr Ile Arg Thr Arg Leu Asn 35 40 45 ata cgc gag tgc tgc gag gatgga tgg tgc tgt act gct gca ccc tta 192 Ile Arg Glu Cys Cys Glu Asp GlyTrp Cys Cys Thr Ala Ala Pro Leu 50 55 60 aca ggt cgt tagggataaaggaaaatggc tttggatgag acccctgcga 241 Thr Gly Arg 65 attgtccctggatgtgagat ttggaaagca gactgttcct ttcgcacgtg ttcgtggaat 301 ttcgaatggtcgttaacaac acgctgccac ttgcaagcca ccatctctct gtcctttcgt 361 atgtggaactgtatgatcta acaactgaaa tgtcagaaag ttttcagtgg gtatacacta 421 tgatcgtata431 31 67 PRT Conus textile 31 Met Arg Cys Phe Pro Val Phe Ile Ile LeuLeu Leu Leu Ile Ala Ser 1 5 10 15 Ala Pro Cys Phe Asp Ala Arg Thr LysThr Asp Asp Asp Val Pro Leu 20 25 30 Ser Ser Leu Arg Asp Asn Leu Lys ArgThr Ile Arg Thr Arg Leu Asn 35 40 45 Ile Arg Glu Cys Cys Glu Asp Gly TrpCys Cys Thr Ala Ala Pro Leu 50 55 60 Thr Gly Arg 65 32 441 DNA Conustextile CDS (1)..(201) 32 atg cgc tgt ttc cca gtc ttc atc att ctt ctgttg cta att gca tct 48 Met Arg Cys Phe Pro Val Phe Ile Ile Leu Leu LeuLeu Ile Ala Ser 1 5 10 15 gca cct tgc ttt gat gcc cga acg aag acc gatgat gat gtg ccc ctg 96 Ala Pro Cys Phe Asp Ala Arg Thr Lys Thr Asp AspAsp Val Pro Leu 20 25 30 tca tct ctc cgc gat aat cta aag cga acg ata cgaaca cgc ctg aac 144 Ser Ser Leu Arg Asp Asn Leu Lys Arg Thr Ile Arg ThrArg Leu Asn 35 40 45 ata cgc ggg tgc tgc gag gat gga tgg tgc tgt act gctgca ccc tta 192 Ile Arg Gly Cys Cys Glu Asp Gly Trp Cys Cys Thr Ala AlaPro Leu 50 55 60 aca ggt cgt tagggataaa ggaaaatggc tttggatgag acccctgcaa241 Thr Gly Arg 65 attgtccctg gatgtgagat ttggaaagca gactgttcctttcgcacgtg ttcgtggaat 301 ttcgaatggt cgttaacaac acgctgccac ttgcaagccaccatctctct gtcctttcgt 361 atgtggaact gtatgatcta acaactgaaa tgtcagaaagttttcagtgg gtatacacta 421 tgatcgtata gtcagtaatt 441 33 67 PRT Conustextile 33 Met Arg Cys Phe Pro Val Phe Ile Ile Leu Leu Leu Leu Ile AlaSer 1 5 10 15 Ala Pro Cys Phe Asp Ala Arg Thr Lys Thr Asp Asp Asp ValPro Leu 20 25 30 Ser Ser Leu Arg Asp Asn Leu Lys Arg Thr Ile Arg Thr ArgLeu Asn 35 40 45 Ile Arg Gly Cys Cys Glu Asp Gly Trp Cys Cys Thr Ala AlaPro Leu 50 55 60 Thr Gly Arg 65 34 416 DNA Conus marmoreus CDS(1)..(210) 34 atg cgc tgc ctc cca gtc ttc gtc att ctt ctg ctg ctg attgca tct 48 Met Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile AlaSer 1 5 10 15 gca cct agc gtt gat gcc cga ccg aag acc aaa gat gat atgccc ctg 96 Ala Pro Ser Val Asp Ala Arg Pro Lys Thr Lys Asp Asp Met ProLeu 20 25 30 gca tct ttc cat gat aat gca aag cga atc ctg caa ata ctt caggac 144 Ala Ser Phe His Asp Asn Ala Lys Arg Ile Leu Gln Ile Leu Gln Asp35 40 45 aga aat ggt tgc tgc aga gca gga gac tgc tgt tca cga ttt gag ata192 Arg Asn Gly Cys Cys Arg Ala Gly Asp Cys Cys Ser Arg Phe Glu Ile 5055 60 aag gaa aat gac ttt gga tgagacccct gcaaactgtc cttggatgtg 240 LysGlu Asn Asp Phe Gly 65 70 agatttggaa agcagactgt tcctttcgca cgtgttcgtggaatttcgaa tggtcgttaa 300 caacacgctg ccacttgcaa gctactatct ctctgtccttttgtctgtgg aactgtatga 360 tcaaacaact gaaatgtcat agaaattttt cagtgggtaaacactatgac catgta 416 35 70 PRT Conus marmoreus 35 Met Arg Cys Leu ProVal Phe Val Ile Leu Leu Leu Leu Ile Ala Ser 1 5 10 15 Ala Pro Ser ValAsp Ala Arg Pro Lys Thr Lys Asp Asp Met Pro Leu 20 25 30 Ala Ser Phe HisAsp Asn Ala Lys Arg Ile Leu Gln Ile Leu Gln Asp 35 40 45 Arg Asn Gly CysCys Arg Ala Gly Asp Cys Cys Ser Arg Phe Glu Ile 50 55 60 Lys Glu Asn AspPhe Gly 65 70 36 487 DNA Conus marmoreus CDS (3)..(179) 36 ga atg cgctgc ctc cca gtc ttc gtc att ctt ctg ctg ctg att gca 47 Met Arg Cys LeuPro Val Phe Val Ile Leu Leu Leu Leu Ile Ala 1 5 10 15 tct gca cct agcgtt gat gcc cga ccg aag acc aaa gat gat atg ccc 95 Ser Ala Pro Ser ValAsp Ala Arg Pro Lys Thr Lys Asp Asp Met Pro 20 25 30 ctg gca tct ttc cacgat aat gca aag cga atc ctg caa ata ctt cag 143 Leu Ala Ser Phe His AspAsn Ala Lys Arg Ile Leu Gln Ile Leu Gln 35 40 45 gac aga aat gct tgc tgcata gta agg cag tgc tgt tgatgatttg 189 Asp Arg Asn Ala Cys Cys Ile ValArg Gln Cys Cys 50 55 agataaagga aaatgacttt ggatgagacc cctgcaaactgtccctggat gtgagatttg 249 gaaagcagac tgttcctttc gcacgtgttc gtggaatttcgaatggtcgt taacaacacg 309 ctgccacttg caagctacta tctctctgtc ctttcatctgtggaactgta tgatcaaaca 369 actgaaatgt catagaaatt tttcagtggg taaacactatgatcatgtag tcagtaatta 429 catcatttgg aattccatca agcttatcga taccgtcgacctcgaggggg ggcccggt 487 37 59 PRT Conus marmoreus 37 Met Arg Cys Leu ProVal Phe Val Ile Leu Leu Leu Leu Ile Ala Ser 1 5 10 15 Ala Pro Ser ValAsp Ala Arg Pro Lys Thr Lys Asp Asp Met Pro Leu 20 25 30 Ala Ser Phe HisAsp Asn Ala Lys Arg Ile Leu Gln Ile Leu Gln Asp 35 40 45 Arg Asn Ala CysCys Ile Val Arg Gln Cys Cys 50 55 38 370 DNA Conus marmoreus CDS(1)..(180) 38 atg cgc tgc ctc cca gtc ttt gtc att ctt ctg ctg ctg attgca tct 48 Met Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu Leu Ile AlaSer 1 5 10 15 gca cct agc gtt gat gcc cga ccg aag acc aaa gat gat atgccc ctg 96 Ala Pro Ser Val Asp Ala Arg Pro Lys Thr Lys Asp Asp Met ProLeu 20 25 30 gca tct ttc cat gat aat gca aag cga atc ctg caa ata ctt caggac 144 Ala Ser Phe His Asp Asn Ala Lys Arg Ile Leu Gln Ile Leu Gln Asp35 40 45 aga aat ggt tgc tgc aga gca gga gac tgc tgt tca tgatttgaga 190Arg Asn Gly Cys Cys Arg Ala Gly Asp Cys Cys Ser 50 55 60 taaagggaaatgactttgga tgagacccct gcaaactgtc cttggatgtg agatttggaa 250 agcagactgttcctttcgca cgtgttcgtg gaatttcgaa tggtcgttaa caacacgctg 310 ccacttgcaagctactatct ctctgtcctt tcatctgtgg aactgtatga tcaaacaact 370 39 60 PRTConus marmoreus 39 Met Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu LeuIle Ala Ser 1 5 10 15 Ala Pro Ser Val Asp Ala Arg Pro Lys Thr Lys AspAsp Met Pro Leu 20 25 30 Ala Ser Phe His Asp Asn Ala Lys Arg Ile Leu GlnIle Leu Gln Asp 35 40 45 Arg Asn Gly Cys Cys Arg Ala Gly Asp Cys Cys Ser50 55 60 40 413 DNA Conus caracteristicus CDS (1)..(192) 40 atg cgc tgtctc ccg gtc ttc atc att ctt ctg ctg ctg att gca tct 48 Met Arg Cys LeuPro Val Phe Ile Ile Leu Leu Leu Leu Ile Ala Ser 1 5 10 15 gca cct ggcgtt gat gcc caa ccg aag acc aaa tat aat gcg ccc ctg 96 Ala Pro Gly ValAsp Ala Gln Pro Lys Thr Lys Tyr Asn Ala Pro Leu 20 25 30 aca tct ctc cacgat aat gca aag ggt ata cta caa gaa cat tgg aac 144 Thr Ser Leu His AspAsn Ala Lys Gly Ile Leu Gln Glu His Trp Asn 35 40 45 aaa cgc tgc tgc cccaga agg ctt gcc tgc tgt att ata gga cgg aaa 192 Lys Arg Cys Cys Pro ArgArg Leu Ala Cys Cys Ile Ile Gly Arg Lys 50 55 60 tgaatgattt tgggtgagatccctgcaaac tgtccctgga tttgaatttt ggaaagcaga 252 ctgttccttt cgcacgtgttcgtggaattt cgaatggtcg ttaacaacac gctgccactt 312 gcaagctact atctctctgtcctttttctc tgtgaaactg gatggtctaa caactgaaat 372 gtcatagaaa attttcaatgggtatactct atgaccatct a 413 41 64 PRT Conus caracteristicus 41 Met ArgCys Leu Pro Val Phe Ile Ile Leu Leu Leu Leu Ile Ala Ser 1 5 10 15 AlaPro Gly Val Asp Ala Gln Pro Lys Thr Lys Tyr Asn Ala Pro Leu 20 25 30 ThrSer Leu His Asp Asn Ala Lys Gly Ile Leu Gln Glu His Trp Asn 35 40 45 LysArg Cys Cys Pro Arg Arg Leu Ala Cys Cys Ile Ile Gly Arg Lys 50 55 60 42410 DNA Conus caracteristicus CDS (1)..(189) 42 atg cgc tgt ctc cca gtcttc atc att ctt ctg ctg ctg att gca tct 48 Met Arg Cys Leu Pro Val PheIle Ile Leu Leu Leu Leu Ile Ala Ser 1 5 10 15 gca cct ggc gtt gat gcccaa ccg aag acc aaa tat gat gcg ccc ctg 96 Ala Pro Gly Val Asp Ala GlnPro Lys Thr Lys Tyr Asp Ala Pro Leu 20 25 30 aca tct ctc cac gat aat gcaaag ggt ata cta caa gaa cat tgg aac 144 Thr Ser Leu His Asp Asn Ala LysGly Ile Leu Gln Glu His Trp Asn 35 40 45 aaa cgc tgc tgc ccc aac aag ccttgc tgt ttt ata gga agg aaa 189 Lys Arg Cys Cys Pro Asn Lys Pro Cys CysPhe Ile Gly Arg Lys 50 55 60 tgaatgattt tgggtgagac ccctgcaaac tgtccctggatttgaatttt ggaaagcaga 249 ctgttccttt cgcacgtgtt cgtggaattt cgaatggtcgttaacaacac gctgccactt 309 gcaagctact atctctctgt cctttttctc tgtgaaactggatggtctaa caactgagat 369 gtcatagaaa attttcaatc ggtgtactct atgaccatct a410 43 63 PRT Conus caracteristicus 43 Met Arg Cys Leu Pro Val Phe IleIle Leu Leu Leu Leu Ile Ala Ser 1 5 10 15 Ala Pro Gly Val Asp Ala GlnPro Lys Thr Lys Tyr Asp Ala Pro Leu 20 25 30 Thr Ser Leu His Asp Asn AlaLys Gly Ile Leu Gln Glu His Trp Asn 35 40 45 Lys Arg Cys Cys Pro Asn LysPro Cys Cys Phe Ile Gly Arg Lys 50 55 60 44 413 DNA Conus quercinus CDS(1)..(186) 44 atg cgc tgt gtc cca gtc ttc atc att ctt ctg ctg ctg agtcca tct 48 Met Arg Cys Val Pro Val Phe Ile Ile Leu Leu Leu Leu Ser ProSer 1 5 10 15 gca cct agc gtt gat gcc cat ccg atg acc aaa gat gat gtaccc cag 96 Ala Pro Ser Val Asp Ala His Pro Met Thr Lys Asp Asp Val ProGln 20 25 30 gca tct ctc cat gat gat gca aag cga acc cta caa gta cct tggatg 144 Ala Ser Leu His Asp Asp Ala Lys Arg Thr Leu Gln Val Pro Trp Met35 40 45 aaa cgc ggg tgc tgc gca atg ttg act tgc tgc gtt gga cga 186 LysArg Gly Cys Cys Ala Met Leu Thr Cys Cys Val Gly Arg 50 55 60 taaagggaaatgactttgga tgagacccct acgaactgtc cctggatgtg aaatttggac 246 agcagactgctcctttcgca cgtgttcgtg gaatttcgaa tggtcgttaa caacacgctg 306 ccacttgcaagctattatct ctctgtccct ttatctgtgg aactggataa tctaacaact 366 gaaacgtcattgaaaatttt caatggatat atattatgat ccatata 413 45 62 PRT Conus quercinus45 Met Arg Cys Val Pro Val Phe Ile Ile Leu Leu Leu Leu Ser Pro Ser 1 510 15 Ala Pro Ser Val Asp Ala His Pro Met Thr Lys Asp Asp Val Pro Gln 2025 30 Ala Ser Leu His Asp Asp Ala Lys Arg Thr Leu Gln Val Pro Trp Met 3540 45 Lys Arg Gly Cys Cys Ala Met Leu Thr Cys Cys Val Gly Arg 50 55 6046 735 DNA Conus gloriamaris CDS (70)..(258) 46 gggcaggtac tcaacgaacttcaggacaca ttcttttcac ctggacacgg gaaactgact 60 ataagcaga atg cgc tac ctacca gtc ttc gtc att ctt ctg ctg ctg att 111 Met Arg Tyr Leu Pro Val PheVal Ile Leu Leu Leu Leu Ile 1 5 10 gca tct ata cct agc gat act gtc caactg aag acc aaa gat gat atg 159 Ala Ser Ile Pro Ser Asp Thr Val Gln LeuLys Thr Lys Asp Asp Met 15 20 25 30 ccc ctg gca tct ttc cac ggt aat ggaaga cga atc ctg cga atg ctt 207 Pro Leu Ala Ser Phe His Gly Asn Gly ArgArg Ile Leu Arg Met Leu 35 40 45 tca aac aaa cgc tta tgc tgt gtc acc gaggat tgg tgc tgt gaa tgg 255 Ser Asn Lys Arg Leu Cys Cys Val Thr Glu AspTrp Cys Cys Glu Trp 50 55 60 tgg taaaggaaaa tgactttgga tgagacccctgcaaactgtt tctggatgtg 308 Trp agatttggaa agcagactgt tctttcgcacgtattcgtga aatttcgaat ggtcgttaac 368 aacacgctgc cacttgcaag ctgctatctctctgtctttt catctgtgga actgtatgat 428 ctaacaactg aaatgtcata gacatttttcattgggtata cactatgacc atgtagccag 488 taattacatc atttggacct tttggatatttttcagtatg taagtgtgtt cccttaaaaa 548 gtcctttgta attatgtatt ttaanaatttangttttgca cataaattgt aaaacgctgt 608 cctttctgtt gntcctacat cantggtggggaaaagnaaa atgtttggcc ntggtcaaat 668 ttaaataatn accctgccgt ttnaatgcngttattantgg tattttnaac nttgnacggt 728 taaactt 735 47 63 PRT Conusgloriamaris 47 Met Arg Tyr Leu Pro Val Phe Val Ile Leu Leu Leu Leu IleAla Ser 1 5 10 15 Ile Pro Ser Asp Thr Val Gln Leu Lys Thr Lys Asp AspMet Pro Leu 20 25 30 Ala Ser Phe His Gly Asn Gly Arg Arg Ile Leu Arg MetLeu Ser Asn 35 40 45 Lys Arg Leu Cys Cys Val Thr Glu Asp Trp Cys Cys GluTrp Trp 50 55 60 48 374 DNA Conus gloriamaris CDS (3)..(188) 48 ga atgcgc tgt ctc cca gtc ttc gtc att ctt ctg ctg ctg att gca 47 Met Arg CysLeu Pro Val Phe Val Ile Leu Leu Leu Leu Ile Ala 1 5 10 15 tct gca cctagc gtt gat gcc caa ccg aag acc aaa gat gat gtg ccc 95 Ser Ala Pro SerVal Asp Ala Gln Pro Lys Thr Lys Asp Asp Val Pro 20 25 30 ctg gca cct ttgcac gat aat ata agg agt act cta caa aca ctt cgg 143 Leu Ala Pro Leu HisAsp Asn Ile Arg Ser Thr Leu Gln Thr Leu Arg 35 40 45 aag aaa gtc tgc tgccgc cca gtg cag gat tgc tgt tca ggg aaa 188 Lys Lys Val Cys Cys Arg ProVal Gln Asp Cys Cys Ser Gly Lys 50 55 60 tgaagggaaa tgaatttggatgagacccct gcgaactgtc cctggatgtg agatttggaa 248 agcagactgt tcctttcgcacgtgttcgtg gaatttcgaa tggtcgttaa caacacgctg 308 ccacttgcaa gctactatctctctgtcctt tcatctgcgg aactggatga cctaaagctt 368 gtgatc 374 49 62 PRTConus gloriamaris 49 Met Arg Cys Leu Pro Val Phe Val Ile Leu Leu Leu LeuIle Ala Ser 1 5 10 15 Ala Pro Ser Val Asp Ala Gln Pro Lys Thr Lys AspAsp Val Pro Leu 20 25 30 Ala Pro Leu His Asp Asn Ile Arg Ser Thr Leu GlnThr Leu Arg Lys 35 40 45 Lys Val Cys Cys Arg Pro Val Gln Asp Cys Cys SerGly Lys 50 55 60

What is claimed is:
 1. A substantially pure τ-conotoxin peptide havingthe generic formula I:Xaa₁-Xaa₂-Xaa₃-Xaa₄-Cys-Cys-Xaa₅-Xaa₆-Xaa₇-Xaa₈-Xaa₉-Cys-Cys-Xaa₁₀-Xaa₁₁-Xaa₁₂-Xaa₁₃-Xaa₁₄-Xaa₁₅-Xaa₁₆-Xaa₁₇-Xaa₁₈-Xaa₁₉(SEQ ID NO:1), wherein Xaa₁ is des-Xaa₁, Asp, Glu or γ-carboxy-Glu(Gla);Xaa₂ is des-Xaa₂, Gln, Asn, Glu, Trp (D or L), neo-Trp, halo-Trp or anyunnatural aromatic amino acid; Xaa₁ is des-Xaa₃, Gly, Ala, Asn or Gln;Xaa₄ is des-Xaa₄,Val, Leu (D or L), Ile, Ala, Gly, Glu, Gla, Asp, Ser,Thr, Phe, Trp (D or L), neo-Trp, halo-Trp (D or L) or any unnaturalaromatic amino acid; Xaa₅ is Pro, hydroxy-Pro, Gln, Asn, Glu, Gla, Ala,Gly, Lys, Arg, Ile, Val, homoarginine, omithine, N-methyl-Lys,N,N-dimethyl-Lys, N,N,N-trimethyl-Lys or any unnatural basic amino acid;Xaa₆ is Val, Phe, Thr, Ser, Glu, Gla, Asp, Asn, Gln, Ala, Gly, Ile, Leu(D or L), Met, Pro, hydroxy-Pro, Arg, homoarginine, ornithine, Lys,N-methyl-Lys, N,N-dimethyl-Lys, N,N,N-trimethyl-Lys, any unnatural basicamino acid or any unnatural aromatic amino acid; Xaa₇ is any Val, Ile,Asn, Leu (D or L), Gln, Gly, Ala, Phe, Glu, Gla, Arg, ornithine,homoarginine, Lys, N-methy-Lys, N,N-dimethyl-Lys, N,N,N-trimethyl-Lys,any unnatural basic amino acid or any unnatural aromatic amino acid;Xaa₈ is Ile, Leu (D or L), Met, Thr, Ser, Pro, hydroxy-Pro, Gln, Asp,Glu, Gla, Asn, Arg, homoarginine, ornithine, Lys, N-methy-Lys,N,N-dimethyl-Lys, N,N,N-trimethyl-Lys, Tyr, nor-Tyr, mono-halo-Tyr,di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr, nitro-Tyr, any unnatural basicamino acid, any unnatural aromatic amino acid or any unnatural hydroxycontaining amino acid; Xaa₉ is des-Xaa₉, Ala, Gly, Asp, Glu, Gla, Trp (Dor L) neo-Trp, halo-Trp (D or L), Lys, N-methy-Lys, N,N-dimethyl-Lys,N,N,N-trimethyl-Lys, Arg, homoarginine, ornithine, Tyr, nor-Tyr,mono-halo-Tyr, di-halo-Tyr, O-sulpho-Tyr, O-phospho-Tyr, nitro-Tyr orany unnatural basic amino acid; Xaa₁₀ is des-Xaa₁₀, Ile, Leu (D or L),Val, Glu, Gla, Asp, Thr, Ser, Pro, hydroxy-Pro, Trp (D or L), neo-Trp,halo-Trp (D or L), Phe, any unnatural aromatic amino acid or anyunnatural hydroxy containing amino acid; Xaa₁₁ is des-Xaa₁₁, Gln, Asn,Leu (D or L), Ile, Val, Ala, Gly, Trp (D or L), neo-Trp, halo-Trp (D orL), Arg, homoarginine, ornithine, Lys, N-methy-Lys, N,N-dimethyl-Lys,N,N,N-trimethyl-Lys, any unnatural basic amino acid or any unnaturalaromatic amino acid; Xaa₁₂ is des-Xaa₁₂, Ala, Gly, Phe, Trp (D or L),neo-Trp, halo-Trp (D or L) or any unnatural aromatic amino acid; Xaa₁₃is des-Xaa₁₃, Glu, Gla, Asp, Phe or any unnatural aromatic amino acid;Xaa₁₄ is des-Xaa₁₄, Ile, Val or Leu (D or L); Xaa₁₅ is des-Xaa₁₅, Thr,Ser, Arg, homoarginine, ornithine, Lys, N-methy-Lys, N,N-dimethyl-Lys,N,N,N-trimethyl-Lys or any unnatural basic amino acid; Xaa₁₆ isdes-Xaa₁₆, Glu, Gla or Asp; Xaa₁₇ is des-Xaa₁₇, Asn or Gln; Xaa₁₈ isdes-Xaa₁₈, Asp, Glu or Gla; Xaa₁₉ is des-Xaa₁₉, Phe or any unnaturalaromatic amino acid; and the C-terminus contains a free carboxyl groupor an amide group.
 2. The substantially pure α-conotoxin peptide ofclaim 1, which is modified to contain an O-glycan, an S-glycan or anN-glycan.
 3. A substantially pure τ-conotoxin peptide consisting ofPhe-Cys-Cys-Xaa₁-Phe-Ile-Arg-Xaa₂-Cys-Cys-Xaa₃ (SEQ ID NO:3); whereinXaa₁ is Pro or hydroxy-Pro; Xaa₂ is Tyr, mono-halo-Tyr, di-halo-Tyr,O-sulpho-Tyr, O-phospho-Tyr or nitro-Tyr; Xaa₃ is Trp or halo-Trp; andthe C-terminus contains a carboxyl or amide group.
 4. The substantiallypure τ-conotoxin peptide of claim 3, wherein Xaa₂ is Tyr.
 5. Thesubstantially pure τ-conotoxin peptide of claim 2, wherein Xaa₃ is Trp.6. The substantially pure τ-conotoxin peptide of claim 2, wherein Xaa₁is Pro or hydroxy-Pro, Xaa₂ is Tyr, mono-iodo-Tyr or di-iodo-Tyr, andXaa₃ is Trp.
 7. The substantially pure τ-conotoxin peptide of claim 2,wherein Xaa₁ is Pro, Xaa₂ is Tyr and Xaa₃ is Trp.
 8. The substantiallypure τ-conotoxin peptide of claim 2 which is modified to contain anO-glycan, an S-glycan or an N-glycan.